KR20200008580A - Use of anti-B7H3 antibodies to treat cancer in the central nervous system - Google Patents

Abstract

The subject matter disclosed herein provides the use of an anti-B7H3 antibody for treating cancer in the CNS, including tumors that are metastatic to the central nervous system (CNS), in particular, meningiocarcinoma.

Description

Use of anti-B7H3 antibodies to treat cancer in the central nervous system

Cross Reference to Related Applications

This application claims priority to US Provisional Application No. 62 / 505,558, filed May 12, 2017, the content of which is incorporated by reference in its entirety and claims priority thereto.

Sequence list

The specification includes a reference to a list of sequences submitted through EFS on May 14, 2018. 37 C.F.R. According to § 1.52 (e) (5), the sequence listing text file named 0727340700SL was 21,716 bytes in size and was created on 14 May 2018. The Sequence Listing, filed electronically on May 14, 2018, does not extend beyond the scope of the specification and therefore does not include new issues.

Field of invention

The subject matter disclosed herein relates to the use of anti-B7H3 antibodies for the treatment of cancers in the CNS, including tumors that are metastatic to the central nervous system (CNS), in particular, meningiocarcinoma.

Adult CNS tumors include primary CNS tumors (formed by cancerous cells that develop within the CNS) and tumors that are metastatic to the CNS (cancer cells that have spread from the primary tumors from other organs in the body to the CNS). About 20,000 new primary CNS tumor cases are diagnosed each year in the United States, with an estimated 24% to 25% of all cancer patients in the United States having brain metastases. The meninges (two linings surrounding the brain and spine) appeared to be an evacuation site that caused relapse. When tumor cells gain access to the cerebrospinal fluid pathway, migrate to distal sites in the brain and spine, settle, and grow, meninge metastasis (LM, also referred to as meningiocarcinoma) occurs. LM has always been widely accepted as fatal.

Primary CNS tumors are the third most common cancer in puberty and young adults (ages 15-39), and are the third leading cause of cancer deaths in this age group. Metastatic CNS tumors, on the other hand, are most common in adults than in children. Thus, there remains a need for innovative treatment for adult CNS tumors.

Summary of the Invention

The subject matter disclosed herein relates to the use of anti-B7H3 antibodies for treating CNS cancers, including primary CNS cancers and cancers that are metastatic to the CNS. In certain embodiments, the anti-B7H3 antibody is administered into the CNS to treat meninge metastasis of cancer in an adult subject.

In certain non-limiting embodiments, the subject matter disclosed herein comprises administering a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds B7H3 into a subject's CNS, for treating cancer in a human subject. Provide a method. In certain embodiments, the cancer is a primary CNS cancer or a cancer that is metastatic to the CNS. In certain embodiments, the antibody or antigen-binding fragment thereof is conjugated to a radioisotope and / or mode of treatment (eg, chelator compound or anticancer agent). In certain embodiments, the human subject is an adult. In certain embodiments, the cancer is metastatic to the meninges. In certain embodiments, the cancer metastatic to the CNS is a solid tumor occurring outside the CNS. In certain embodiments, the solid tumor is selected from the group consisting of sarcoma, melanoma, ovarian cancer and rhabdomyosarcoma. In certain embodiments, the solid tumor is selected from the group consisting of melanoma, ovarian cancer and rhabdomyosarcoma. In certain embodiments, the central nervous system (CNS) cancer is selected from the group consisting of neuroblastoma and primary recurrent CNS malignancy. In certain embodiments, the cancer metastatic to the CNS is a solid tumor selected from the group consisting of breast cancer (eg, triple-negative breast cancer) and lung cancer (eg, small cell lung cancer and non-small cell lung cancer). In certain embodiments, the antibody or antigen-binding fragment is 8H9, derived from 8H9, and / or structurally related to 8H9, including but not limited to murine, humanized, chimeric, and human versions of 8H9 (see below). .

In certain non-limiting embodiments, the subject matter disclosed herein is a method for administering to a subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof conjugated to a radioisotope and / or other therapeutic modality that specifically binds human B7H3. Provided is a method of treating cancer in a human subject, comprising the step. In certain embodiments, the cancer is any cancer, including B7H3 positive cancers or tumor cells. In certain embodiments, the cancer is primary or metastatic to the subject's CNS and the antibody is administered into the subject's CNS. In certain embodiments, the subject is an adult. In certain embodiments, the subject is not an adult.

In certain non-limiting embodiments, the antibody or antigen-binding fragment thereof that specifically binds human B7H3 is a murine antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a chimeric antibody or antigen-binding fragment thereof And human antibodies or antigen-binding fragments thereof. In certain embodiments, the antibody is a murine antibody or antigen-binding fragment thereof. In certain embodiments, the antibody or antigen-binding fragment thereof binds to the FG-loop of B7H3. In certain embodiments, the antibody or antigen-binding fragment is 8H9, derived from 8H9, and / or structurally related to 8H9, including but not limited to murine, humanized, chimeric, and human versions of 8H9 (see below). .

In certain embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3 (NYDIN), (b) the amino acid sequence shown in SEQ ID NO: 4 (WIFPGDGSTQY) Heavy chain variable region CDR2, (c) heavy chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5 (QTTATWFAY), (d) light chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6 (RASQSISDYLH), (e) Light chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 7 (YASQSIS), and / or (f) light chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 8 (QNGHSFPLT). In certain embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 1, and / or (b) a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 2. Include.

In certain embodiments, the antibody or antigen-binding fragment thereof comprises at least (a) a heavy chain variable region CDR comprising the amino acid sequence shown in SEQ ID NO: 3 (NYDIN), SEQ ID NO: 4 (WIFPGDGSTQY), or SEQ ID NO: 5 (QTTATWFAY) and (b) a light chain variable region CDR comprising the amino acid sequence shown in SEQ ID NO: 6 (RASQSISDYLH), SEQ ID NO: 7 (YASQSIS), or SEQ ID NO: 8 (QNGHSFPLT).

In certain embodiments, the antibody or antigen-binding fragment thereof is administered intradurally to a subject. In certain embodiments, the antibody or antigen-binding fragment thereof is administered to a subject via an intraventricular device. In certain embodiments, the intraventricular device is an intraventricular catheter. In certain embodiments, the ventricular device is an intraventricular reservoir.

In certain embodiments, the radioisotope conjugated with the antibody or fragment is ¹²⁴ I, ¹³¹ I, ¹⁷⁷ Lu, or ^99m Tc.

In certain embodiments, the methods disclosed herein further comprise administering to the subject an antibody or antigen-binding fragment thereof in one treatment cycle. In certain embodiments, the method comprises administering two treatment cycles of the antibody or antigen-binding fragment thereof to the subject. In certain embodiments, one treatment cycle comprises a dosimetric dose and a therapeutic dose. In certain embodiments, the therapeutically effective amount is about 10 mCi to about 200 mCi. In certain embodiments, the therapeutically effective amount is about 50 mCi.

In certain embodiments, the method extends the survival of the subject compared to a control subject or control subject population that has not received treatment. In certain embodiments, the method extends sedation of cancer in a subject as compared to a control subject or control subject population that has not received treatment.

In certain embodiments, the antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least about 80%, about 90%, about 95%, about 99% or about 100% homology with the amino acid sequence set forth in SEQ ID NO: 17. . In certain embodiments, the antibody or antigen-binding fragment thereof comprises the amino acid sequence set forth in SEQ ID NO: 17. In certain embodiments, the antibody or antigen-binding fragment thereof has amino acids 224-241 of SEQ ID NO: 17. In certain embodiments, the antibody or antigen-binding fragment thereof has amino acids 242-267 of SEQ ID NO: 17.

In certain embodiments, the treatment modality is selected from the group consisting of one or more chelator compounds, one or more chemotherapeutic agents, one or more checkpoint inhibitory agents, and radiation therapy. In certain embodiments, a therapeutic mode other than a radioisotope is conjugated to the antibody. In certain embodiments, the mode of treatment is a chelator compound. In certain embodiments, the radioisotope is indirectly bound to the antibody or antigen-binding fragment thereof via a chelator compound such as DOTA, DOTA-like compound, or DTPA. In certain embodiments, the antibody or antigen-binding fragment thereof is conjugated to a chelator compound, wherein the chelator compound is bound to a radioisotope. In certain embodiments, the chelator compound is DOTA or DTPA. In certain embodiments, the antibody or antigen-binding fragment thereof is conjugated to a chelator compound (eg DOTA, DTPA, or related compound) and the chelator is a radioisotope (eg, ¹²⁴ I, ¹³¹ I, ¹⁷⁷ Lu, or ^99m Tc). ) In vitro or in vivo.

In certain embodiments, the mode of treatment is monoclonal antibody 3F8 (MoAb 3F8), granulocyte-macrophage-colon-stimulating factor (GM-CSF), or a combination thereof. In certain embodiments, the mode of treatment is administered systemically to the subject's CNS and / or to the subject. In certain embodiments, the mode of treatment is administered to a subject simultaneously or sequentially with the antibody or antigen-binding fragment thereof.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds human B7H3, wherein the antibody or antigen-binding fragment thereof is conjugated to a chelator compound and the chelator compound is radioisotope Is bound to an element.

In certain embodiments, the antibody is selected from the group consisting of murine antibodies and antigen-binding fragments thereof, humanized antibodies and antigen-binding fragments thereof, chimeric antibodies and antigen-binding fragments thereof, and human antibodies and antigen-binding fragments thereof. In certain embodiments, the antibody or antigen-binding fragment thereof is a murine antibody or antigen-binding fragment thereof. In certain embodiments, the antibody or antigen-binding fragment thereof binds to the FG-loop of B7H3.

In certain embodiments, the antibody or antigen-binding fragment thereof comprises a) a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3, b) a heavy chain variable region CDR2, c) sequence comprising the amino acid sequence shown in SEQ ID NO: 4 Heavy chain variable region CDR3 comprising the amino acid sequence shown in No. 5, d) light chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6, e) light chain variable region CDR2 and f) comprising the amino acid sequence shown in SEQ ID NO: 7 Light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 8.

In certain embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1 and (b) a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2.

In certain embodiments, the radioisotope is ¹²⁴ I, ¹³¹ I, ¹⁷⁷ Lu, or ⁹⁹ mTc. In certain embodiments, the chelator compound is DOTA or DTPA.

In certain embodiments, the antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least about 80%, about 90%, about 95%, about 99% or about 100% homology with the amino acid sequence set forth in SEQ ID NO: 17. . In certain embodiments, the antibody or antigen-binding fragment thereof comprises the amino acid sequence set forth in SEQ ID NO: 17. In certain embodiments, the antibody or antigen-binding fragment thereof has amino acids 224-241 of SEQ ID NO: 17. In certain embodiments, the antibody or antigen-binding fragment thereof has amino acids 242-267 of SEQ ID NO: 17.

In certain embodiments, the mode of treatment is monoclonal antibody 3F8 (MoAb 3F8), granulocyte-macrophage-colon-stimulating factor (GM-CSF), or a combination thereof. In certain embodiments, the mode of treatment is administered systemically to the subject's CNS and / or to the subject. In certain embodiments, the mode of treatment is administered to a subject simultaneously or sequentially with the antibody or antigen-binding fragment thereof.

In certain embodiments, the antibody or antigen-binding fragment thereof is a DOTA-8H9 conjugate or DTPA-8H9 conjugate. In certain embodiments, the antibody or antigen-binding fragment thereof is a ¹⁷⁷ Lu-DOTA-8H9 conjugate or ¹⁷⁷ Lu-DTPA-8H9 conjugate or (177) LU-CHX-A "-DTPA-8H9.

In certain embodiments, the antibody or antigen-binding fragment thereof is a single chain variable fragment (scFv). In certain embodiments, the scFv comprises a portion of the amino acid sequence set forth in SEQ ID NO: 9, SEQ ID NO: 13, and SEQ ID NO: 14.

In another aspect, the present disclosure provides a composition comprising an antibody or antigen-binding fragment thereof disclosed herein.

In another aspect, the present disclosure provides a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof disclosed herein and a pharmaceutically acceptable carrier.

In another aspect, the present disclosure provides a method of imaging a tumor in a subject comprising administering an antibody or antigen-binding fragment thereof disclosed herein to the subject.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof disclosed herein for use as a medicament.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof disclosed herein for use in the treatment of cancer.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof disclosed herein for use in the treatment of central nervous system (CNS) cancer.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof disclosed herein for use in the treatment of metastatic CNS neuroblastoma, sarcoma, melanoma, ovarian carcinoma and primary recurrent CNS malignancy.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof disclosed herein for use in a method of imaging a tumor in a subject.

In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof disclosed herein for use in the methods disclosed herein.

In another aspect, the present disclosure provides the use of an antibody or antigen-binding fragment thereof disclosed herein for the manufacture of a medicament for tumor cell contrast carrying an antigen recognized by the antibody or antigen-binding fragment thereof.

In another aspect, the present disclosure provides the use of an antibody, or antigen-binding fragment thereof, disclosed herein for the manufacture of a medicament for the method disclosed herein.

A. In certain non-limiting embodiments, the subject matter disclosed herein comprises administering a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds human B7H3 into the subject's CNS. Provided are methods for treating CNS cancers, wherein the cancer is a primary central nervous system (CNS) cancer or a cancer metastatic to the CNS, and the antibody or fragment is conjugated to a radioisotope and / or other mode of treatment.

A1. The method of A, wherein the cancer is metastatic to the meninges.

A2. The method of A, wherein the cancer metastatic to the CNS is a non-CNS solid tumor.

A3. The method of A2, wherein the solid tumor is selected from the group consisting of melanoma, ovarian cancer and rhabdomyosarcoma.

A4. The method of A, wherein the antibody is selected from the group consisting of murine antibodies, humanized antibodies, chimeric antibodies and human antibodies.

A5. The method of A, wherein the antibody is a murine antibody.

A6. The method of A, wherein the antibody or antigen-binding fragment thereof binds to the FG-loop of B7H3.

A7. In A, the antibody or antigen-binding fragment thereof is

(a) a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3 (NYDIN),

(b) a heavy chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 4 (WIFPGDGSTQY),

(c) a heavy chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5 (QTTATWFAY),

(d) a light chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6 (RASQSISDYLH),

(e) a light chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 7 (YASQSIS) and

(f) light chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 8 (QNGHSFPLT)

How to include.

A8. In A, the antibody or antigen-binding fragment thereof is

(a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1, and

(b) a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2

How to include.

A9. The method of A, wherein the antibody or antigen-binding fragment thereof is administered intradurally to the subject.

A10. The method of A, wherein the antibody or antigen-binding fragment thereof is administered to the subject via an intraventricular device.

A11. The method of A, wherein the intraventricular device is an intraventricular catheter.

A12. The method of A, wherein the ventricular device is an intraventricular reservoir.

A13. The method of A, wherein the radioisotope is ¹³¹ I, ¹⁷⁷ Lu, or ^99m Tc.

A14. The method of A, comprising administering to the subject an antibody or antigen-binding fragment thereof in one treatment cycle.

A15. The method of A, comprising administering two treatment cycles of the antibody or antigen-binding fragment thereof to the subject.

A16. The method of A, wherein one treatment cycle comprises a dosimetric dose and a therapeutic dose.

A17. The method of A, wherein the therapeutically effective amount is about 10 mCi to about 200 mCi or about 10 mCi to about 100 mCi.

A18. The method of A, wherein the therapeutically effective amount is about 50 mCi.

A19. The method of A, which extends the survival of the subject.

A20. The human of A, wherein the antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least about 80%, about 90%, about 95%, about 99% or about 100% homology with the amino acid sequence set forth in SEQ ID NO: 17. Binding to B7H3 Polypeptides.

A21. The method of A, wherein the antibody or antigen-binding fragment thereof binds to a human B7H3 polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 17.

A22. The method of A, wherein the antibody or antigen-binding fragment thereof binds to a human B7H3 polypeptide having amino acids 224-241 of SEQ ID NO: 17.

A23. The method of A, wherein the antibody or antigen-binding fragment thereof binds to a human B7H3 polypeptide having amino acids 242-267 of SEQ ID NO: 17.

A24. The method of A, further comprising administering to the subject a further mode of treatment, such as, but not limited to, one or more chemotherapeutic agents, one or more checkpoint inhibitory agents, and / or radiation therapy. Such one or more additional therapeutic agents may be administered CNS and / or systemically concurrently or sequentially with the B7H3-derived antibody or antigen-binding fragment described herein.

Figure 1. Survival of pediatric neuroblastoma patients treated with MSK prior to cRIT 8H9 compared to all ¹³¹ I-8H9-treated patients.
Figure 2. Survival of pediatric neuroblastoma patients treated with MSK before 2003 compared to ¹³¹ I-8H9-treated patients. 64 patients treated with full CNS induction therapy (blue line), 29 patients treated with ¹³¹ I-8H9 and other therapies (red line) and 19 patients treated with MSK prior to protocol initiation (purple line) in 2003. Survival was compared by Kaplan-Meier analysis.
3. Age based survival upon initial neuroblastoma diagnosis.
4A-4B. Time to initial radiological improvement in groups of patients (4a and 4b) with measurable disease at study entry. The length of the horizontal bar is equal to the duration of subject involvement in protocol 03-133 or post-study follow-up. Radiologic improvement was determined by comparison of pre-treatment scan and post-treatment scan. Improvement is defined as complete response, partial response, or no evidence of disease. Diamond is the first radiological improvement date after the first cycle of 131I-8H9. The white right arrow indicates patients who survived their condition. ▼ The black down arrow indicates the date of death. The Y axis represents individual patients and the X axis represents time. Y = radioactive response, N = no radioactive response mentioned.
5. Multipathic topical CNS neuroblastoma that is calm and over 7 years old. CNS recurrence demonstrating numerous on, under, and spinal metastases.
6. Subgroup analysis and effects on overall patient survival. The effect of certain variables on overall patient survival was assessed by Kaplan-Meier analysis of subgroups of patients treated with ¹³¹ I-8H9. (a) The effect of age on survival was assessed in patients under 18 months of age (blue line) and patients over 18 months of age (red line) at the time of initial neuroblastoma diagnosis. (b) The effect of MYCN status on survival was evaluated in ¹³¹ I-8H9-treated patients with amplified MYCN tumors (blue line) and patients with unamplified MYCN tumors (red line). (c) The effect of the age of registration in the protocol on survival was assessed in patients enrolled in the protocol in 2003-2009 (blue line) and patients enrolled in 2010-2016 (red line). (d) The effect of prior CSI therapy on survival was assessed in patients not treated with CSI prior to ¹³¹ I-8H9 cRIT (blue line) and in patients treated with CSI (red line).

All publications, patents, and other references cited herein are hereby incorporated by reference in their entirety.

For purposes of clarity and not of limitation, the description is divided into the following subsections:

1. Definition

2. Anti-B7H3 Antibody

3. Treatment Method

1. Definition

In the description that follows, certain conventions will be followed regarding the use of the term. In general, terms used herein are to be interpreted consistently with the meaning of these terms as they are known to those skilled in the art.

As used herein, the term “antibody” refers to fragments of antibody molecules that retain immunogen-binding capacity as well as intact antibody molecules. Such fragments are also well known in the art and are routinely adopted both in vitro and in vivo. Thus, as used herein, the term "antibody" refers to intact immunoglobulin molecules as well as the well-known active fragments F (ab ') _{2 and} Fab. Fab fragments without F (ab ') _{2 and} Fc fragments of intact antibodies are more quickly removed from circulation and may have less non-specific tissue binding of intact antibodies (Wahl et al., J. Nucl. Med . 24: 316-325 (1983). Antibodies of the invention include whole intact antibodies, bispecific antibodies; Chimeric antibodies; Fab, Fab ', single chain V region fragment (scFv), fusion polypeptides and unusual antibodies. In certain embodiments, the antibody is a glycoprotein comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as V _H ) and a heavy chain constant (C _H ) region. The heavy chain constant region consists of three domains, CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated herein as V _L ) and a light chain constant C _L region. The light chain constant region consists of one domain, C _L. The V _H and V _L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FR). Each V _H and V _L consists of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody may mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (C1 q) of the traditional complement system.

The terms “antigen-binding portion”, “antigen-binding fragment”, or “antigen-binding region” of an antibody, as used interchangeably herein, refer to a region or portion of an antibody that binds to an antigen and imparts antigen specificity to the antibody. Represent; Antigen-binding proteins, such as fragments of antibodies, include one or more fragments of an antibody that retain specific binding capacity to an antigen (eg, peptide / HLA complex). It has been shown that the antigen-binding function of antibodies can be performed by fragments of full length antibodies. Examples of antigen-binding portions encompassed within the term “antibody fragment” of an antibody include Fab fragments that are monovalent fragments consisting of V _L , V _H , C _L and CH1 domains; F (ab) ₂ fragment, which is a bivalent fragment comprising two Fab fragments linked by disulfide bridges in the hinge region; Fd fragment consisting of the V _H and CH1 domains; An Fv fragment consisting of the V _L and V _H domains of one arm of the antibody; V _H DAb fragment consisting of domains (Ward et al., 1989 Nature 341: 544-546); And an isolated complementarity determining region (CDR).

"CDR" is defined as the complementarity determining region amino acid sequence of an antibody which is the hypervariable region of an immunoglobulin heavy and light chain. See, eg, Kabat et al., Sequences of Proteins of Immunological Interest, 4th U.S. Department of Health and Human Services, National Institutes of Health (1987). As used herein, the term “high variable region” or “HVR” refers to a sequence that is highly variable (“complementarity determining region” or “CDR”) and / or to forming a structurally defined loop (“high variable loop”) and And / or each region of an antibody variable domain containing an antigen-contacting residue (“antigen contact”). In general, an antibody comprises three heavy chains and three light chain CDRs or CDR regions in the variable region. The CDRs provide most of the contact residues for binding of the antibody to the antigen or epitope.

In addition, the two domains of the Fv fragment, V _L and V _H , are encoded by separate genes, but they are produced by a synthetic linker that allows the V _L and V _H regions to be paired to form a single protein chain that forms a monovalent molecule. It can be linked using recombinant methods. These are known as single chain Fv (scFv); See, eg, Bird et al., 1988 Science 242: 423-426; And Huston et al., 1988 Proc. Natl. Acad. Sci. 85: 5879-5883. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as intact antibodies.

As used herein, an antibody that "specifically binds to B7H3" is 5 × 10 ^-7 M or less, 1 × 10 ^-7 M or less, 5 × 10 ^-8 M or less, 1 × 10 ^-8 M or less, 5 × 10 ^-9 M or less, 1 × 10 ^-9 M or less, 5 × 10 ^-10 M or less, 1 × 10 ^-10 M or less, 5 × 10 ^-11 M or less, or 1 × 10 ^-11 B7H3 having a K _d of less than M (Eg, human B7H3) is shown.

An antigen such as an antibody that competes for binding with a reference antibody for binding to B7H3 or an antibody that cross-competites for binding is about binding of the reference antibody to an antigen (eg, B7H3) in a competition assay. At least 50%, such as at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% At least about 98% or at least about 99%, whereas the reference antibody is at least about 50%, such as at least about 55%, at least about 60, in the competition assay for binding of the antibody to the antigen (eg, B7H3). Block at least%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% . Exemplary competition assays are described in "Antibodies," Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY) (1988). In certain embodiments, the reference antibody is a murine anti-B7H3 antibody. In certain embodiments, the reference antibody is 8H9.

An antigen, such as an antibody or antigen-binding fragment that competes for binding with a reference antibody for binding to B7H3 or an antibody or antigen-binding fragment that cross-competites for binding, refers to an antigen (eg, Binding of the reference antibody to B7H3) at least about 50%, such as at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85% , At least about 90%, at least about 95%, at least about 98% or at least about 99% of the antibody or antigen-binding fragment, whereas the reference antibody is the antibody or antigen against the antigen (eg, B7H3) in a competition assay. At least about 50%, such as at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90 Block at least%, at least about 95%, at least about 98% or at least about 99%. Exemplary competition assays are described in "Antibodies," Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY) (1988).

Sequence homology or sequence identity is typically described by sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or It is measured using the PILEUP / PRETTYBOX program. Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. In an exemplary approach for determining the degree of identity, the BLAST program can be used, with sequences of probability between ^{e -3} and ^e-100 indicating closely related sequences.

An “therapeutically effective amount” of an agent, such as an anti-B7H3 antibody or antigen-binding fragment thereof, may be used to achieve the desired therapeutic or prophylactic result, such as primary cancer for the CNS or metastatic cancer for the CNS, such as the meninges. Effective dose during the dosage and period of time necessary to treat).

As used herein, a “subject” can be a human or non-human subject such as but not limited to non-human primates, dogs, cats, horses, rodents, rabbits, and the like. An adult human subject is a subject at least 18 years old or at least 20 years old. Adult non-human subjects are those who have achieved sexual maturity. Human subjects who are not adults are pediatric subjects.

As used herein, "administering into a subject's CNS" means administration to one or more of the subject's cerebrospinal fluid, subarachnoid space, meningoid tissue, and / or nervous (brain and / or spinal) tissue.

As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to clinical intervention in an attempt to alter the natural course of an individual being treated and for prevention or during the course of clinical pathology. Can be performed. Desired therapeutic effects include, but are not limited to, prolongation of survival, prevention of disease recurrence, alleviation of symptoms, weakening of any direct or indirect pathological consequences of the disease, prevention of metastasis, slowing of disease progression, alleviation or improvement of disease state, and sedation or improvement Included prognosis. In certain embodiments, antibodies of the subject matter disclosed herein are used to delay the development of a disease or to slow the progression of a disease, such as cancer primary to the CNS or metastatic to the CNS (eg, meninges).

As used herein, the term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i. will be. For example, "about" can mean within 3 or more than 3 standard deviations, depending on the practice in the art. Alternatively, "about" can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, more preferably up to 1% of a given value . Alternatively, especially for biological systems or processes, the term may mean within 10-fold, preferably within 5-fold, more preferably within 2-fold of the value.

Any concentration range, percentage range, ratio range, or integer range described herein includes any integer value within the range recited and, where appropriate, fractions thereof (e.g., 1/10 and 1/100 of an integer) unless otherwise indicated. It is understood that.

2. Anti-B7H3 Antibody

The subject matter disclosed herein is the use of an anti-B7H3 antibody or antigen-binding fragment thereof for treating a cancer, such as a cancer that is primary to the CNS or metastatic to the CNS (eg, to the parenchyma or to a soft membrane). To provide. The anti-B7H3 antibody may be a murine, humanized, chimeric, or human antibody.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof binds the B7H3 polypeptide. In certain embodiments, the B7H3 polypeptide is a human B7H3 polypeptide. The B7H3 polypeptide comprises at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% of SEQ ID NO: 17, or a fragment thereof, as provided below, May have an amino acid sequence that is about 90%, about 95%, about 99%, or about 100% homologous (homologous herein may be determined using standard software such as BLAST or FASTA) and / or optionally up to one Or up to two or up to three amino acid substitutions (eg, conservative substitutions). In certain embodiments, the B7H3 polypeptide comprises the amino acid sequence shown in SEQ ID NO: 17. In certain embodiments, the B7H3 polypeptide has at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 150, at least 200 , At least 250, at least 300, at least 350, at least 400, at least 450, at least 500 and at most 534 amino acids in length. Alternatively or additionally, in various non-limiting embodiments, the B7H3 polypeptide may comprise amino acids 1-534, 1-50, 50-100, 100-150, 150-200, 200-250, 224 of SEQ ID NO: 17. Have an amino acid sequence of from 241, 242 to 267, 241 to 267, 250 to 300, 300 to 350, or 350 to 400, 400 to 450, 450 to 500, and 500 to 534. In certain embodiments, the B7H3 polypeptide comprises amino acids 224 to 241 of SEQ ID NO: 17. Amino acids 224 to 241 of SEQ ID NO: 17 have an amino acid sequence of NPVLQQDAHSSVTITPQR (SEQ ID NO: 15). In certain embodiments, the B7H3 polypeptide has amino acids 242 to 267 of SEQ ID NO: 17. Amino acids 242 to 267 of SEQ ID NO: 17 have an amino acid sequence of SPTGAVEVQVPEDPVVALVGTDATLR (SEQ ID NO: 16).

MLRRRGSPGMGVHVGAALGALWFCLTGALEVQVPEDPVVALVGTDATLCCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFAEGQDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYQGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSILRVVLGANGTYSCLVRNPVLQQDAHSSVTITPQRSPTGAVEVQVPEDPVVALVGTDATLRCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFTEGRDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYRGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSVLRVVLGANGTYSCLVRNPVLQQDAHGSVTITGQPMTFPPEALWVTVGLSVCLIALLVALAFVCWRKIKQSCEEENAGAEDQDGEGEGSKTALQPLKHSDSKEDDGQEIA (SEQ ID NO: 17)

In certain embodiments, the anti-B7H3 antibody is a murine antibody. In certain embodiments, the anti-B7H3 antibodies are all antibody 8H9 disclosed in US Pat. Nos. 7,737,258, 7,666,424, 8,148,154, 7,740,845, 8,414,892, 9,062,110 and 8,501,471 and International Patent Publication No. WO2008 / 116219, which are incorporated by reference in their entirety.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3 (NYDIN), a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 4 (WIFPGDGSTQY) CDR2, heavy chain variable region CDR3 comprising amino acid sequence (QTTATWFAY) as shown in SEQ ID NO: 5, light chain variable region CDR1 comprising amino acid sequence (RASQSISDYLH) as shown in SEQ ID NO: 6, amino acid sequence as shown in SEQ ID NO: 7 (YASQSIS) Light chain variable region CDR2, and / or light chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 8 (QNGHSFPLT). In certain embodiments, the anti-B7H3 antibody comprises (a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1, and / or (b) a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2. SEQ ID NOs 1-8 are provided below.

QVQLQQSGAELVKPGASVKLSCKASGYTFTNYDINWVRQRPEQGLEWIGWIFPGDGSTQYNEKFKGKATLTTDTSSSTAYMQLSRLTSEDSAVYFCARQTTATWFAYWGQGTLVTVSAAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK (SEQ ID NO: 1)

DIVMTQSPATLSVTPGDRVSLSCRASQSISDYLHWYQQKSHESPRLLIKYASQSISGIPSRFSGSGSGSDFTLSINSVEPEDVGVYYCQNGHSFPLTFGAGTKLELKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLTKNSDSEKSE

NYDIN (SEQ ID NO: 3)

WIFPGDGSTQY (SEQ ID NO: 4)

QTTATWFAY (SEQ ID NO: 5)

RASQSISDYLH (SEQ ID NO: 6)

YASQSIS (SEQ ID NO: 7)

QNGHSFPLT (SEQ ID NO: 8)

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof cross-compete for binding to antibodies 8H9 and B7H3. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3 (NYDIN), a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 4 (WIFPGDGSTQY) CDR2, heavy chain variable region CDR3 comprising amino acid sequence (QTTATWFAY) as shown in SEQ ID NO: 5, light chain variable region CDR1 comprising amino acid sequence (RASQSISDYLH) as shown in SEQ ID NO: 6, amino acid sequence as shown in SEQ ID NO: 7 (YASQSIS) And cross-compete for binding to B7H3 with a reference antibody comprising the light chain variable region CDR2 and the light chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 8 (QNGHSFPLT). In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1 and (b) a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2. Cross-compete for binding to the reference antibody and B7H3.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof binds to the same epitope on antibodies 8H9 and B7H3. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3 (NYDIN), a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 4 (WIFPGDGSTQY) CDR2, heavy chain variable region CDR3 comprising amino acid sequence (QTTATWFAY) as shown in SEQ ID NO: 5, light chain variable region CDR1 comprising amino acid sequence (RASQSISDYLH) as shown in SEQ ID NO: 6, amino acid sequence as shown in SEQ ID NO: 7 (YASQSIS) To a reference antibody comprising a light chain variable region CDR2 and a light chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 8 (QNGHSFPLT) and the same epitope on B7H3. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1 and (b) a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2. It binds to the same reference antibody and the same epitope on B7H3.

In certain embodiments, the anti-B7H3 antibody is a single chain variable fragment (scFv). The scFv can be murine, humanized or human scFv. In certain embodiments, the anti-B7H3 antibody is a murine scFv. In certain embodiments, the anti-B7H3 antibody is an scFv comprising the amino acid sequence shown in SEQ ID NO: 9 (provided below). In certain embodiments, the anti-B7H3 antibody is an scFv comprising the amino acid sequence shown in SEQ ID NO: 13 (provided below). In certain embodiments, the anti-B7H3 antibody is an scFv comprising the amino acid sequence shown in SEQ ID NO: 14 (provided below).

QVKLQQSGAELVKPGASVKLSCKASGYTFTNYDINWVRQRPEQGLEWIGWIFPGDGSTQYNEKFKGKATLTTDTSSSTAYMQLSRLTSEDSAVYFCARQTTATWFAYWGQGTTVTVSSGGGGSGGGGSGGGGSDIELTQSPTTLSVTPGDRVSLSCRASQSISDYLHWYQQKSHESPRLLIKYASQSISGIPSRFSGSGSGSDFTLSINSVEPEDVGVYYCQNGHSFPLTFGAGTKLELKQAA (SEQ ID NO: 9)

QVKLQQSGAELVKPGASVKLSCKASGYTFTNYDINWVRQRPEQGLEWIGWIFPGDGSTQYNEKFKGKATLTTDTSSSTAYMQLSRLTSEDSAVYFCARQTTATWFAYWGQGTTVTVSSDGGGSGGGGSGGGGSDIELTQSPTTLSVTPGDRVSLSCRASQSISDYLHWYQQKSHESPRLLIKYASQSISGIPSRFSGSGSGSDFTLSINSVEPEDVGVYYCQNGHSFPLTFGAGTKLELKQAA (SEQ ID NO: 13)

QVKLQQSGAELVEPGASVKLSCKASGYTFTNYDINWVRQRPEQGLEWIGWIFPGDGSTQYNEKFKGKATLTTDTSSSTAYMQLSRLTSEDSAVYFCARQTTATWFAYWGQGTTVTVSSDGGGSGGGGSGGGGSDIELTQSPTTLSVTPGDQVSLSCRASQSISDYLHWYQQKSHESPQLLIKYASQSISGIPSRFSGSGSGSDFTLSINSVEPEDVGVYYCQNGHSFPLTFGAGTELELEQAA (SEQ ID NO: 14)

The nucleotide sequences encoding SEQ ID NO: 9 are SEQ ID NO: 10 (sense) and SEQ ID NO: 11 (complementary chain). SEQ ID NOs: 10 and 11 are provided below.

caggtcaaac tgcagcagtc tggggctgaa ctggtaaagc ctggggcttc agtgaaattg tcctgcaagg cttctggcta caccttcaca aactatgata taaactgggt gaggcagagg cctgaacagg gacttgagtg gattggatgg atttttcctg gagatggtag tactcaatac aatgagaagt tcaagggcaa ggccacactg actacagaca catcctccag cacagcctac atgcagctca gcaggctgac atctgaggac tctgctgtct atttctgtgc aagacagact acggctacct ggtttgctta ctggggccaa gggaccacgg tcaccgtctc ctcaggtgga ggcggttcag gcggaggtgg ctctggcggt ggcggatcgg acatcgagct cactcagtct ccaaccaccc tgtctgtgac tccaggagat agagtctctc tttcctgcag ggccagccag agtattagcg actacttaca ctggtaccaa caaaaatcac atgagtctcc aaggcttctc atcaaatatg cttcccaatc catctctggg atcccctcca ggttcagtgg cagtggatca gggtcagatt tcactctcag tatcaacagt gtggaacctg aagatgttgg agtgtattac tgtcaaaatt gtcacactgctctctctctct

gtccagtttg acgtcgtcag accccgactt gaccatttcg gaccccgaag tcactttaac aggacgttcc gaagaccgat gtggaagtgt ttgatactat atttgaccca ctccgtctcc ggacttgtcc ctgaactcac ctaacctacc taaaaaggac ctctaccatc atgagttatg ttactcttca agttcccgtt ccggtgtgac tgatgtctgt gtaggaggtc gtgtcggatg tacgtcgagt cgtccgactg tagactcctg agacgacaga taaagacacg ttctgtctga tgccgatgga ccaaacgaat gaccccggtt ccctggtgcc agtggcagag gagtccacct ccgccaagtc cgcctccacc gagaccgcca ccgcctagcc tgtagctcga gtgagtcaga ggttggtggg acagacactg aggtcctcta tctcagagag aaaggacgtc ccggtcggtc tcataatcgc tgatgaatgt gaccatggtt gtttttagtg tactcagagg ttccgaagag tagtttatac gaagggttag gtagagaccc taggggaggt ccaagtcacc gtcacctagt cccagtctaa agtgagagtc atagttgtca caccttggac ttctacaacc tcacataatg acagttttac cagtgtcgcc aggcgagtga ccgcc

In certain embodiments, the nucleotide encoding the scFv that binds to the B7H3 polypeptide has the nucleotide sequence shown in SEQ ID NO: 12 (provided below).

caggtcaaac tgcagcagtc tggggctgaa ctggtaaagc ctggggcttc agtgaaattg tcctgcaagg cttctggcta caccttcaca aactatgata taaactgggt gaggcagagg cctgaacagg gacttgagtg gattggatgg atttttcctg gagatggtag tactcaatac aatgagaagt tcaagggcaa ggccacactg actacagaca catcctccag cacagcctac atgcagctca gcaggctgac atctgaggac tctgctgtct atttctgtgc aagacagact acggctacct ggtttgctta ctggggccaa gggaccacgg tcaccgtctc ctcagatgga ggcggttcag gcggaggtgg ctctggcggt ggcggatcgg acatcgagct cactcagtct ccaaccaccc tgtctgtgac tccaggagat agagtctctc tttcctgcag ggccagccag agtattagcg actacttaca cctggtaccaa caaaaatcac atgagtctcc aaggcttctc atcaaatatg cttcccaatc catctctggg atcccctcca ggttcagtgg cagtggatca gggtcagatt tcactctcag tatcaacagt gtggaacctg aagatgttgg agtgtattac tgtcaaaatt gccacactgctct

In certain embodiments, the anti-B7H3 antibody is a humanized antibody. In certain embodiments, the anti-B7H3 antibody is a humanized anti-B7H3 antibody disclosed in International Patent Publication No. WO2016 / 033225, which is incorporated by reference in its entirety. In certain embodiments, both anti-B7H3 antibodies are humanized B7-H3-reactive antibodies disclosed in US Pat. Nos. 8,802,091 and 9,441,049, which are incorporated by reference in their entirety.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof cross-competes for binding to B7H3 with a humanized anti-B7H3 antibody disclosed in International Patent Publication No. WO2016 / 033225. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof binds to the same epitope on B7H3 as the humanized anti-B7H3 antibody disclosed in International Patent Publication No. WO2016 / 033225.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof cross-competes for binding to B7H3 with humanized B7-H3-reactive antibodies disclosed in US Pat. Nos. 8,802,091 and 9,441,049. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof binds to the same epitope on B7H3 as the humanized B7-H3-reactive antibody disclosed in US Pat. Nos. 8,802,091 and 9,441,049.

For example, and not by way of limitation, cross-competitive antibodies are antibody 8H9, which is a humanized anti-B7H3 antibody disclosed in International Patent Publication No. WO2016 / 033225, or humanized B7 disclosed in US Pat. Nos. 8,802,091 and 9,441,049. -H3-reactive antibody) and the same epitope region, such as the same epitope, adjacent epitopes or overlapping epitopes.

Such cross-competitive antibodies can be identified based on their ability to cross compete with reference antibodies in standard B7H3 binding assays. For example, Biacore assays, ELISA assays or flow cytometry can be used to demonstrate cross-competition with reference antibodies. The ability of an assay antibody to inhibit binding of a reference antibody to B7H3 (eg, human B7H3) allows the assay antibody to compete with the reference antibody for binding to B7H3 and thus to bind the same epitope region on the reference antibody and B7H3. Demonstrate In certain embodiments, the cross-competitive antibody binds to the same epitope on the reference antibody and B7H3 (eg, human B7H3).

As a non-limiting example of a competition assay, an immobilized antigen such as a B7H3 (eg, human B7H3) polypeptide is evaluated for a first labeled antibody that binds to the antigen and its ability to compete with the first antibody for binding to the antigen. It may be incubated in a solution comprising the recipient second unlabeled antibody. In certain embodiments, the second antibody may be present in the hybridoma supernatant. As a control, the immobilized antigen is incubated in a solution comprising the first labeled antibody rather than the second unlabeled antibody. After incubation under conditions acceptable for binding of the first antibody to the antigen, excess unbound antibody is removed and the amount of label associated with the immobilized antigen is measured. If the amount of label associated with the immobilized antigen in the evaluation sample relative to the control sample is substantially reduced, such as above about 50%, this suggests that the second antibody is competing with the first antibody for binding to the antigen. . See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch. 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

In certain embodiments, the cross-competitive antibody is about 5 × 10 ⁻⁷ M or less, about 1 × 10 ⁻⁷ M or less, about 5 × 10 ⁻⁸ M or less, about 1 × 10 with respect to B7H3 (eg, human B7H3) K _d of ^{8 −8} M or less, about 5 × 10 ⁻⁹ M or less, about 1 × 10 ⁻⁹ M or less, about 5 × 10 ⁻¹⁰ M or less, or about 1 × 10 ⁻¹⁰ M or less.

In certain embodiments, the anti-B7H3 antibody is conjugated to a radioisotope to produce a cytotoxic radiopharmaceutical, also referred to as a radioimmune conjugate. Examples of radioisotopes that can be conjugated to antibodies for diagnostic or therapeutic use include, but are not limited to, ²¹¹ At, ¹⁴ C, ⁵¹ Cr, ⁵⁷ Co, ⁵⁸ Co, ⁶⁷ Cu, ¹⁵² Eu, ⁶⁷ Ga, ³ H, ¹¹¹ In, ⁵⁹ Fe, ²¹² Pb, ¹⁷⁷ Lu, ³² P, ²²³ Ra, ²²⁴ Ra, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁷⁵ Se, ³⁵ S, ^99m Tc, ²²⁷ Th, ⁸⁹ Zr, ⁹⁰ Y, ¹²³ I, ¹²⁴ I, ¹²⁵ I, ¹³¹ I and alpha-emitting particles are included. Non-limiting examples of alpha-emitting particles include ²⁰⁹ Bi, ²¹¹ Bi, ²¹² Bi, ²¹³ Bi, ²¹⁰ Po, ²¹¹ Po, ²¹² Po, ²¹⁴ Po, ²¹⁵ Po, ²¹⁶ Po, ²¹⁸ Po, ²¹¹ At, ²¹⁵ At, ²¹⁷ At , ²¹⁸ At, ²¹⁸ Rn, ²¹⁹ Rn, ²²⁰ Rn, ²²² Rn, ²²⁶ Rn, ²²¹ Fr, ²²³ Ra, ²²⁴ Ra, ²²⁶ Ra, ²²⁵ Ac, ²²⁷ Ac, ²²⁷ Th, ²²⁸ Th, ²²⁹ Th, ²³⁰ Th, ²³² Th, ²³¹ Pa, ²³³ U, ²³⁴ U, ²³⁵ U, ²³⁶ U, ²³⁸ U, ²³⁷ Np, ²³⁸ Pu, ²³⁹ Pu, ²⁴⁰ Pu, ²⁴⁴ Pu, ²⁴¹ Am, ²⁴⁴ Cm, ²⁴⁵ Cm, ²⁴⁸ Cm, ²⁴⁹ Cf and ²⁵² Cf. In certain embodiments, the radioisotope can be selected from ^{94 m} Tc, ⁶⁴ Cu, ⁶⁸ Ga, ⁶⁶ Ga, ⁷⁶ Br, ⁸⁶ Y, ⁸² Rb, ^{110 m} In, ¹³ N, ¹¹ C, and ¹⁸ F. Methods of making radioimmune conjugates are established in the art. Examples of radioimmunoconjugates including Zevalin ™ (IDEC Pharmaceuticals) and Bexxar ™ (Corixa Pharmaceuticals) are commercially available, and similar methods can be used to prepare radioimmunoconjugates using the antibodies of the invention.

Radiolabeled antibody preparations can be prepared according to techniques well known in the art. For example, monoclonal antibodies can be iodinated by contact with sodium and / or potassium iodide and chemical oxidizing agents such as sodium hypochlorite, or enzymatic oxidizing agents such as lactoperoxidase. Provided antibody preparations are labeled with technetium-99m by ligand exchange methods, for example by reducing pertechate to stannous solution, chelating the reduced technetium on a Sephadex column, and applying the antibody to the column. Can be. In certain embodiments, provided antibody formulations are labeled using direct labeling techniques, such as by incubating the antibody and buffer solutions such as pertechate, reducing agents such as SNCI2, sodium-potassium phthalate solution. Intermediate functional groups used to bind to radioisotopes, often present as metal ions for antibodies, are diethylenetriaminepentaacetic acid (DTPA), or ethylene diaminetetraacetic acid (EDTA), or 1,4,7,10-tetra Azacyclododecane-l, 4,7,10-tetraacetic acid (DOTA), or p-aminobenzyl-DOTA (DOTA-Bn). Radioactive isotopes can be detected, for example, by dosimetry.

1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (also known as DOTA) is an organic compound having the formula (CH ₂ CH ₂ NCH ₂ CO ₂ H) ₄ . This molecule consists of a central 12-membered tetraaza ring (ie containing four nitrogen atoms). DOTA is used as a complexing agent, especially for lanthanide ions. The complex has medical use as contrast agent and cancer treatment. Several forms of DOTA exist, each with different kinetics and stability constants. Bifunctional chelating agents can bind to metal and still retain chemically reactive functional groups for covalent attachment to the peptide.

DOTA can be conjugated to monoclonal antibodies by attachment of one of four carboxyl groups as amides.

Pentetic acid or diethylenetriaminepentaacetic acid (DTPA) is an aminopolycarboxylic acid consisting of a diethylenetriamine backbone having five carboxymethyl groups. DTPA is represented by the molecular formula C ₁₄ H ₂₃ N ₃ O ₁₀ and IUPAC name 2- [bis [2- [bis (carboxymethyl) amino] ethyl] amino] acetic acid. DTPA is an edetate and chelating agent used in the manufacture of radiopharmaceuticals. DTPA can improve tumor cell radiocytotoxicity by chelating metal moieties of unbound, extracellular radioimmunotherapy to higher levels of local aggregation of radioimmunotherapy, while leaving normal tissues free from radiocytotoxic effects. DTPA is also used in radiographic procedures as a complex with radioisotopes. As a chelating agent, DTPA forms up to eight bonds to wrap around metal ions. Since transition metals usually form less than eight coordination bonds, DTPA still has binding capacity to other reagents after complexing with these metals.

In certain embodiments, the number of DOTA molecules or DTPA molecules used for conjugation per antibody formulation is comprised of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 DOTA molecules or DTPA molecules. Selected from the group. In certain embodiments, DOTA or DTPA is an intermediate functional group. In certain embodiments, the radiolabeled antibody comprises two or three DOTA molecules or DTPA molecules.

High specific activity is achievable when used with DOTA-peptides radiolabeled with ⁹⁰ Y, ¹¹¹ In, ⁶⁸ Ga and ¹⁷⁷ Lu. Higher inactivation can be achieved when radiolabeling DTPA-peptides. While DTPA-peptides can be radiolabeled at room temperature, the main disadvantage of using DOTA-peptides is that the introduction of radiometals requires heat and time. Although a certain amount of radioactivity is required for the detection of target tissue uptake by the imaging system, consistently low mass doses of DTPA-peptides can be administered.

In certain embodiments, the radiolabeled antibody formulation is a ¹⁷⁷ Lu-DOTA-8H9 conjugate or a ¹⁷⁷ Lu-DTPA-8H9 conjugate or (177) LU-CHX-A "-DTPA-8H9.

In certain embodiments, the pediatric human subject is administered to a CNS by, but not limited to, administration of a therapeutically effective amount of ¹⁷⁷ Lu-DOTA-8H9, ¹⁷⁷ Lu-DTPA-8H9 or (177) LU-CHX-A "-DTPA-8H9. In certain related embodiments, ¹⁷⁷ Lu-DOTA-8H9, ¹⁷⁷ Lu-DTPA-8H9 or (177) LU-CHX-A "-DTPA-8H9 are pediatric humans. Administered into the subject's CNS.

3. Treatment Method

An anti-B7H3 antibody of the subject matter disclosed herein is a human subject suffering from cancer (eg, a cancer primary to the CNS, or a cancer metastatic to the CNS) in an amount sufficient to prevent, inhibit, or reduce progression. Eg, for therapeutic treatment on an adult human subject). Progression includes, for example, cancer growth, invasion, metastasis and / or relapse.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof prolongs the survival of a subject compared to a control subject or control subject population that has not received the treatment. In certain embodiments, the survival period is extended at least about 25%, or at least about 30%, or at least about 50%.

In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof extends the sedation of a cancer of a subject relative to a control subject or control subject population that has not received the treatment.

 In certain embodiments, the method comprises administering to the subject a therapeutically effective amount of an anti-B7H3 antibody or antigen-binding fragment thereof (or a pharmaceutical composition thereof) to produce an anticancer effect in the subject. In certain embodiments, an "anticancer effect" is defined as a decrease in aggregate cancer cell mass, a decrease in cancer cell growth rate, a decrease in cancer cell proliferation, a decrease in tumor mass, a decrease in tumor volume, a decrease in cancer cell proliferation, a rate of cancer growth rate. Reduction or reduction of cancer metastasis. In certain embodiments, the anticancer effect is a reduction in cancer cell number. In certain embodiments, where the cancer is a solid tumor, the anticancer effect may be a decrease in tumor size and / or a decrease in tumor growth rate. In certain embodiments, the anticancer effect is a reduction of NB cells in cerebrospinal fluid. In certain embodiments, the anticancer effect is a reduction in aggregate cancer cell burden. In certain embodiments, the anticancer effect is a decrease in the rate of cell proliferation and / or an increase in the rate of cell death. In certain embodiments, the anticancer effect is prolongation of survival. In certain embodiments, the anticancer effect is an extension of the interval until relapse compared to a control subject or control subject population that has not received the treatment.

The therapeutically effective amount may depend on the severity of the disease and the general state of the subject's own immune system. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof is conjugated to a radioisotope, such as those disclosed herein (eg, ¹³¹ I). In certain embodiments, the therapeutically effective amount is about 1 mCi to about 200 mCi, such as about 1 mCi to about 10 mCi, about 10 mCi to about 200 mCi, about 10 mCi to about 160 mCi, about 10 mCi to about 120 mCi, About 10 mCi to about 100 mCi, about 10 mCi to about 70 mCi, about 10 mCi to about 50 mCi, about 50 mCi to about 200 mCi, about 100 to about 200 mCi, about 100 mCi to about 120 mCi, about 120 mCi To about 160 mCi, about 50 mCi to about 100 mCi, about 40 mCi to about 60 mCi, about 60 mCi to about 80 mCi, or about 80 mCi to about 100 mCi. In certain embodiments, the therapeutically effective amount is about 40 mCi to about 60 mCi. In certain embodiments, the therapeutically effective amount is about 10 mCi, about 20 mCi, about 30 mCi, about 40 mCi, about 50 mCi, about 60 mCi, about 70 mCi, about 80 mCi, about 90 mCi, about 100 mCi, about 110 mCi, about 120 mCi, about 130 mCi, about 140 mCi, about 150 mCi, about 160 mCi, about 170 mCi, about 180 mCi, about 190 mCi, or about 200 mCi. In certain embodiments, the therapeutically effective amount is about 50 mCi. In certain embodiments, the therapeutically effective amount is about 100 mCi or less. In certain embodiments, the therapeutically effective amount is about 50 mCi or less.

In certain embodiments, the method comprises administering to the subject an anti-B7H3 antibody or antigen-binding fragment thereof in one treatment cycle. In certain embodiments, the method comprises administering to the subject up to 2, up to 3, up to 4, or up to 5 treatment cycles of the anti-B7H3 antibody or antigen-binding fragment thereof. In certain embodiments, the method comprises administering to the subject two treatment cycles of the anti-B7H3 antibody or antigen-binding fragment thereof. In certain embodiments, the method comprises administering an additional treatment cycle (two new treatment cycles) to the relapsed patient.

In certain embodiments, one treatment cycle comprises a therapeutic dose. In certain embodiments, the therapeutic dose is about 1 mCi to about 100 mCi, such as about 1 mCi to about 10 mCi, about 10 mCi to about 100 mCi, about 10 mCi to about 40 mCi, about 10 mCi to about 70 mCi, About 10 mCi to about 50 mCi, about 50 mCi to about 100 mCi, about 40 mCi to about 60 mCi, about 60 mCi to about 80 mCi, or about 80 mCi to about 100 mCi. In certain embodiments, the therapeutic dose is about 40 mCi to about 60 mCi. In certain embodiments, the therapeutic dose is about 50 mCi. In certain embodiments, the therapeutic dose is administered during the first week of one treatment cycle. In certain embodiments, the therapeutic dose is administered during the second week of one treatment cycle.

In certain embodiments, one treatment cycle comprises a dosimetric dose and a therapeutic dose. In certain embodiments, the dosimetric dose is about 1 mCi to about 10 mCi, such as about 1 mCi to about 3 mCi, about 3 mCi to about 5 mCi, about 5 mCi to about 7 mCi, or about 7 mCi to about 10 mCi). In certain embodiments, the dosimetric dose is about 1 mCi, about 2 mCi, about 3 mCi, about 4 mCi, about 5 mCi, about 6 mCi, about 7 mCi, about 8 mCi, about 9 mCi, or about 10 mCi. In certain embodiments, the dosimetric dose is about 2 mCi. In certain embodiments, the dosimetric dose is administered prior to the therapeutic dose. In certain embodiments, the dosimetric dose is administered during the first week of one treatment cycle.

In certain embodiments, one treatment cycle further comprises an observation period. In certain embodiments, the observation period lasts about 2 weeks, followed by the treatment dose. In certain embodiments, one treatment cycle further comprises a post-treatment assessment. In certain embodiments, the post-treatment assessment lasts about 1 week, following the observation period.

In certain embodiments, the treatment is administered after the subject has been treated with one or more other cancer treatments. In certain embodiments, the treatment is administered simultaneously or sequentially while the subject is being treated with one or more other cancer treatments. Examples of such other cancer treatments include, but are not limited to surgery, chemotherapy, checkpoint inhibitors and radiation.

In certain embodiments, the subject has not exhibited any objective disease progression (eg, determined by neurological or radiological examination) after treatment propensity in the first treatment cycle (eg, after about 4 weeks of treatment dose in the first treatment cycle). A second treatment cycle is administered to the subject if no third or fourth adverse event (eg, as defined by the National Cancer Institute (NCI)) has been experienced. Grade 3 adverse events are generally defined as "severe and undesirable adverse events (hospital or invasive interventions; transfusion; electrolyte intervention radiological procedures; major symptoms requiring treatment endoscopy or surgery)". Class 4 adverse events generally refer to "life-threatening or disturbing adverse events (acute, life-threatening metabolic or cardiovascular complications such as circulation failure, bleeding, sepsis as complications. Life-threatening physiological consequences) ; Intensive care or emergency invasive procedures; necessity for emergency interventional radiotherapy, therapeutic endoscopy or surgery). Controllable fever, headache, nausea and vomiting are not unexpected Grade 3 or 4 adverse events.

In certain embodiments, the subject has up to about 0.5 mg, up to about 1 mg, up to about 2 mg, up to about 3 mg, up to about 4 mg, up to about 5 mg, up to about 6 mg, up to about 7 mg, Up to about 8 mg, up to about 9 mg, up to about 10 mg, up to about 15 mg, or up to about 20 mg of an anti-B7H3 antibody. In certain embodiments, the subject receives at least about 0.5 mg, at least about 1 mg, at least about 2 mg, at least about 3 mg, at least about 4 mg, or at least about 5 mg of anti-B7H3 antibody per treatment cycle.

In certain embodiments, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, or about 10 weeks. Lasts for a while. In certain embodiments, one treatment cycle lasts about 5 weeks.

In certain embodiments, the volume of cerebrospinal fluid equal to the volume of anti-B7H3 antibody to be injected is removed prior to administration of the anti-B7H3 antibody. In certain embodiments, the injection rate does not exceed 1 ml / min during administration of the anti-B7H3 antibody.

The dosing schedule will also vary with the disease state and the subject's condition, typically from a single bolus dose per day or from continuous infusion to multiple doses, or as indicated by the condition of the practitioner and subject.

Identification of medical conditions treatable by anti-B7H3 antibodies is certainly within the skill and knowledge of those skilled in the art. For example, human subjects with primary CNS cancers or cancers metastatic to the CNS are suitable for administration of anti-B7H3 antibodies. A clinician in the art can easily determine whether an individual is a candidate for such treatment, eg, by clinical assessment, physical examination, and the use of medical history / family history. In certain embodiments, the cancer is metastatic to the meninges. In certain embodiments, the cancer is a solid tumor.

Non-limiting examples of primary CNS cancers that can be treated with anti-B7H3 antibodies include pineal blastoma, ventricular melanoma, medulloblastoma, chordoma, astrocytoma, glioblastoma, atypical malformed hepatic tumor (ATRT), multilayer rosette-containing embryonic tumor (ETMR ) And portal plexus carcinoma. In certain embodiments, the cancer is selected from the group consisting of pineal blastoma, ventricular blastoma, medulloblastoma, chordoma, astrocytoma and glioblastoma. In certain embodiments, the cancer is a solid tumor, for example, pineal blastoma, ventricular stromal tumor, medulloblastoma, astrocytoma, glioblastoma or chordoma.

Non-limiting examples of cancers that are metastatic to the CNS (eg, metastatic to the meninges) that can be treated with an anti-B7H3 antibody include sarcomas, retinoblastoma, melanoma, ovarian cancer, rhabdomyosarcoma, breast cancer, and lung cancer (eg small cell). Lung cancer (SCLC) and non-small cell lung cancer (NSCLC)). In certain embodiments, the cancer is selected from the group consisting of melanoma, ovarian cancer, rhabdomyosarcoma, breast cancer, and lung cancer (eg, small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC)). In certain embodiments, the cancer is melanoma. In certain embodiments, the cancer is ovarian cancer.

In certain embodiments, the cancer that can be treated with an anti-B7H3 antibody is an 8H9 reactive cancer. 8H9 reactive cancers are disclosed in US Patent Publication No. 2002/0102264, which is incorporated by reference in its entirety. 8H9 reactive cancers include, but are not limited to, cancers of various lineages: neuroectodermal, mesenchymal and epithelial.

Any suitable method or route may be used to administer the anti-B7H3 antibody or antigen-binding fragment thereof into the CNS. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof is administered intradurally. In certain embodiments, the anti-B7H3 antibody or antigen-binding fragment thereof is administered via an intraventricular device. In certain embodiments, the intraventricular device is an intraventricular catheter. In certain embodiments, the ventricular device is an intraventricular reservoir, such as but not limited to an Ommaya reservoir. In certain embodiments, administration can be performed by the spinal tap or intraparenchy.

The anti-B7H3 antibody can be administered in the form of a composition further comprising a pharmaceutically acceptable carrier. Suitable pharmaceutically acceptable carriers include, for example, one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and the like, as well as combinations thereof. Pharmaceutically acceptable carriers may further comprise small amounts of accessory ingredients, such as hydrating or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the binding protein. Injectable compositions can be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal, as is well known in the art.

In certain embodiments, the subject is administered an additional mode of treatment. Non-limiting examples of treatment modalities include chemotherapeutic agents, checkpoint inhibitory agents, and radiation therapy. In certain embodiments, the mode of treatment is monoclonal antibody 3F8 (MoAb 3F8), granulocyte-macrophage-colon-stimulating factor (GM-CSF), or a combination thereof. Such one or more additional therapeutic agents may be administered into the CNS and / or systemically concurrently or sequentially with the B7H3-derived antibody or antigen-binding fragment described herein.

Example

The subject matter disclosed herein will be better understood with reference to the following examples, which are provided as illustrations of the subject matter disclosed herein rather than as a limitation.

Example 1: ¹³¹ A method of treating neuroblastoma metastatic to the central nervous system using I-8H9.

background

Neuroblastomas (CNS NBs) that are metastatic to the central nervous system are difficult to treat and almost consistently fatal (less than 6 months of median survival and less than 10% of 36 months survival). Intraventricular compartmental radioimmunotherapy (cRIT) with radio-iodinated monoclonal antibody ¹³¹ I-8H9 provides a therapeutic strategy for eradicating NB cells in cerebrospinal fluid.

Clinical studies were conducted to demonstrate the clinical effectiveness of ¹³¹ I-8H9 cRIT for prolonging survival of CNS NB subjects.

Study design and treatment protocol

Eligible subjects at the Memorial Sloan Kettering Cancer Center (MSK) received radiological or pathological confirmation of the CNS NB. Enrolled subjects underwent (1) MSK temozolamide / irinotecan-based CNS rescue therapy, including ¹³¹ I-8H9 cRIT and systemic immunotherapy, or (2) non-therapy therapy with ¹³¹ I-8H9 cRIT. Data is shown as overall survival after diagnosis of CNS metastasis.

One treatment cycle with ¹³¹ I-8H9 was as follows:

Week 1: ¹³¹ I-8H9 (Dose measurement dose: 2 mCi contrast assessment dose)

Week 2: ¹³¹ I-8H9 (therapeutic dose)

Weeks 3 and 4: Observation period

Week 5: repeated MRI of the head and spine, cerebrospinal fluid cytology

Subjects received up to two cycles of ¹³¹ I-8H9 therapy. To assess cumulative toxicity and pharmacokinetics and to evaluate systemic immune responses (eg, human anti-mouse antibodies) to ¹³¹ I-8H9, there was no objective disease progression (neurologic or radiological) 4 weeks after the last intraventricular injection. Subjects without unexpected grade 3 or 4 toxicity (not including controllable fever or headache, nausea, vomiting) were eligible for a second injection at the same dose administered during the first course. Subjects underwent the same treatment plan and post-treatment assessment after the second treatment injection.

result

Of the 105 subjects with CNS NB admitted to MSK since protocol initiation, 80 were treated with ¹³¹ I-8H9 cRIT, including 57 who completed complete CNS specific therapy. The median age of 80 subjects treated with ¹³¹ I-8H9 cRIT was 4.39 years old.

Study start doses ranged from 10 mCi ¹³¹ I-8H9 per cycle for each subject and dose levels ranged up to 80 mCi ¹³¹ I-8H9. In particular, 80 subjects with neuroblastoma CNS / LM metastases received doses of 10 mCi to 70 mCi. The specific activity was on average approximately 5 mCi / mg ¹³¹ I-8H9 in the 10-50 mCi dose range and approximately 50 mCi / mg ¹³¹ I-8H9 in the 50-100 mCi dose range. Subjects under 3 years of age had their doses adjusted.

Of 19 subjects with evaluable cancer, treatment with ¹³¹ I-8H9 caused at least partial response in 7 (36%) subjects. In the analysis, 45 (56%) of ¹³¹ I-8H9-treated subjects, including 36 (45%) who survived at least 36 months and 23 (29%) who survived at least 60 months, were still between 4.8 and 4.8 after CNS metastases. Survival was 152 months (median 58 months). In contrast, the past population of 19 CNS NB subjects treated in MSK before cRIT became standard care in the laboratory (1989-2003) was 2 days after CNS metastasis, including 2 (11%) who survived at least 36 months. Lived from 44 months (median 5.5 months) and no survivors exceeded 44 months. Subgroup analysis of ¹³¹ I-8H9-treated subjects identified age (less than 18 months) and ubiquity of recurrent disease (isolated only by the CNS) at initial NB diagnosis as a positively correlated factor with survival; Amplification of the MYCN oncogene, timing of study enrollment, and complementary cerebral spine investigations were not factors associated with survival in these subjects.

conclusion

76% of subjects with CNS NB treated in MSK received ¹³¹ I-8H9 cRIT and approximately half completed aggressive CNS rescue therapy with ¹³¹ I-8H9 cRIT. Despite advanced CNS involvement, including multiple parenchymal substances with or without meninge disease in 42% of subjects, more than 50% of subjects treated with ¹³¹ I-8H9 cRIT are still alive and nearly 50% at least 36 months Survived. ¹³¹ I-8H9 cRIT represents a significant advance in the treatment of CNS NB, which currently lacks satisfactory standard therapy and is almost consistently fatal.

Example 2: For Central Nervous System / Mental Peel (CNS / LM) Tumors in Adult Subjects ¹³¹ Intradural radioimmunotherapy with I-8H9.

Thirteen adult subjects over 18 years old and with CNS / LM tumors were treated with ¹³¹ I-8H9 using cRIT. The mean age of these 13 adult subjects, ranging from 19.4 years to 53.5 years, was 35.1 years old. The diagnosis of 13 treated adult subjects included primary CNS tumors (pineal blastoma N = 1, ventricular blastoma N = 2, medulloblastoma N = 3, chordoma N = 1) and tumors metastatic to CNS (melanoma N = 3). Rhabdomyosarcoma N = 2 and ovarian cancer N = 1).

Two of 13 subjects after dosimetric dosing were excluded from the study due to progressive disease and compliance concerns. The remaining 11 adult subjects were treated with ¹³¹ I-8H9 cRIT according to the protocol disclosed in Example 1 at a therapeutic dose of 10 mCi to 80 mCi. Five of 11 subjects received a second treatment dose following the second cycle of ¹³¹ I-8H9 therapy. Overall, the average dose received by 11 subjects was about 50 mCi and the total dose ranged from about 10 to about 160 mCi.

MRI review after the first treatment dose included radiological improvement in two subjects, both with metastatic rhabdomyosarcoma; Stable disease in three subjects; Five subjects showed progressive disease; One patient did not have a disease at entry of the protocol. ^The average survival time after ¹³¹ I-8H9 cRIT was greater than 1 year. ^Of the 11 adult subjects who received ¹³¹ I-8H9 cRIT treatment, one subject survived 92 months, 1 survived 17 months, and 2 survived about 6 months after the first injection of ¹³¹ I-8H9.

Although the subject matter disclosed herein has been described in some detail by way of illustration and as an example for purposes of clarity of understanding, the description and examples should not be construed as limiting the scope of the subject matter disclosed herein. The disclosures of all patent and scientific literature cited herein are expressly incorporated by reference in their entirety.

Example  3: In patients with B7-H3 expressing central nervous system malignancies Ventricle  To 131I-8H9 Radioimmune Therapy : 1/2 phase  Research

Introduction

Several tumors have a known propensity for the development of central nervous system (CNS) metastases. Radiation remains a difficult challenge, and despite the aggressive treatment (1, 2), the prognosis remains dark. Brain infiltration and deposition of radiolabeled tumor-specific monoclonal antibodies after convective enhanced delivery or compartmental intraventricular administration (cRIT) can overcome disorders of the blood brain barrier and improve detection and treatment (3-10). . Phase 1 clinical studies at the Memorial Sloan Kettering Cancer Center (MSK) demonstrated the validity of cRIT using anti-GD2 murine mAb 3F8 labeled with iodine-131 (11).

8H9 is a murine mAb specific for B7-H3, a surface immunoregulatory glycoprotein distributed on the cell membranes of several pediatric and adult solid tumors (12). When radiolabeled with iodine-124, 8H9 can be used to assess drug localization and dosimetry using positron emission tomography (PET). When radiolabeled with iodine-131, 8H9 can deliver therapeutic doses of radiation and suppress tumors in mice (13). Iodine-131 penetrates up to 3 mm and releases beta-radiation which leads to DNA damage and binding and cell death of nearby tumor cells and tumor vasculature. cRIT

To improve the dark prognosis of primary and metastatic tumors of the central nervous system (CNS), compartmental radioimmunotherapy (cRIT) with intraventricular ¹²⁴ I- and ¹³¹ I-labeled 8H9 targeting B7-H3 in a phase 1/2 clinical study. Was administered.

To improve the dark prognosis of primary and metastatic tumors of the central nervous system (CNS), compartmental radioimmunotherapy (cRIT) with intraventricular ¹²⁴ I- and ¹³¹ I-labeled 8H9 targeting B7-H3 in a Phase 1/2 clinical study. ) Was administered.

Patient and method

Briefly, eligibility criteria included B7-H3 reactive CNS primary or metastatic tumors, appropriate cerebrospinal fluid (CSF) flow, major organ toxicity below grade 3, platelet 50,000 / μL and ANC 1000 / μL. Patients were awarded ¹²⁴ I- or ¹³¹ I-8H9 of 2 mCi intraventricular for imaging and ¹³¹ I-8H9 of 10- to -80 mCi for treatment. Dosimetry was based on continuous CSF and blood sampling and scintigraphy over 48 hours. Follow-up magnetic resonance imaging was performed at week 5. Injections were repeated in the absence of grade 3 or 4 toxic or progressive disease. Tumor response and overall survival (OS) were noted.

Patients with recurrent primary or metastatic CNS tumors were enrolled in the MSK protocol (clinicalstudys.gov NCT00089245) assessing ¹³¹ I-8H9 cRIT (2004-2017). 8H9 was purified and radiolabeled in MSK using the iodogen technique (14). Inactivation averaged about 5 mCi / mg ¹³¹ I-8H9 in the 2- to 40 mCi dose range, and about 50 mCi / mg ¹³¹ I-8H9 for therapeutic doses of 50 mCi or higher.

Informed consent was obtained from the patient or guardian. Eligibility criteria include B7-H3 reactivity of tumors by immunohistochemistry, stable neurological status, no obstructive or symptomatic hydrocephalus, absolute neutrophil count above 1000 / μL, platelet count above 50,000 / μL, serum bilirubin below 3.0 mg / dl And less than 2 mg / dl serum creatinine. Prior local or cerebral spinal cord (CSI) or chemotherapy was allowed, but not within 3 weeks of enrollment. Indwelling ventricular access device (eg, Ommaya catheter) location, opening, and CSF flow were assessed by a pre-treated 111-indium diethylene triamine pentaacetic acid (DTPA) study.

Clinical and Disease Assessment

Pre- and post-treatment assessments included detailed history, physical examination, total blood count (CBC), comprehensive profile, thyroid function assessment and CSF for total protein, glucose, cell count, cytology. All patients underwent baseline magnetic resonance imaging (MRI) studies of the brain and spine with and without gadolinium within 3 weeks and 1 month before cRIT. In neuroblastoma patients, stage evaluation was performed according to the International Neuroblastoma Stage Evaluation System (15, 16). CNS neuroblastoma is defined as meningoid disease or metastatic deposition in the CNS parenchyma or dura, with the exception of skull-based metastases; Disease assessments for neuroblastoma patients also included ¹²³ I-metho-iodobenzylguanidine (MIBG), computed tomography of the primary site (CT), and bone marrow inhalation and biopsy. After completion of cRIT ¹³¹ I-8H9, disease status was assessed by MRI and CSF cytology of the brain and spine approximately every three months and included CT scans, MIBG scans, and bone marrow assessment of primary sites in neuroblastoma patients.

Dosimetry estimate

Patients were given contrast dose 2 mCi ¹²⁴ I-8H9 or ¹³¹ I-8H9, followed by CSF and blood sampling, and three PET or SPECT scans over 48 hours to assess the dosimetry prior to treatment dosing. The distribution and active concentration of radioactivity on the cerebrospinal axis and the radiation dose to disease plaques and surrounding normal tissues were determined. ¹³¹ I-8H9 radiation exposure of CSF, ventricle, spine, normal brain and blood was based on the assumption of full local absorption of ¹³¹ I beta radiation. The aliquots measured were counted to estimate time-dependent activity concentrations. Each time-activity data was fitted to the exponential function and integrated to yield an area under the curve (AUC) that is attenuated and exhibited cumulative activity concentrations in blood and CSF as described above (11).

¹³¹ I-8H9 treatment

In order to minimize thyroid uptake and to prevent possible meningitis reactions, patients were predosed with oral SSKI drops, Cytomel and acetaminophen before injection. Dexamethasone was administered as 1 mg (0.5 mg for patients less than 20 kg) twice daily for 6 doses, starting from the evening before injection.

The starting dose of this period was chosen to be 10 mCi ¹³¹ I-8H9 based on preclinical studies, and phase 2 studies of 10 mCi ¹³¹ I-3F8 (maximum total 40 mCi) injections can be safely administered 24 (24). Patients had 10- to -80 mCi ¹³¹ I-8H9, 5 mCi / mg at dose levels of 1--4; Single doses of 50 mCi / mg were given at dose levels of 5--8. Dose escalation was performed for every 3 to 6 patients by 10 mCi. Since 2009, a flat therapy dose of 50 mCi ¹³¹ I-8H9 has been administered to the expansion cohort, and myelosuppression is expected in the multi-pre-treatment population at doses above 50 mCi. Toxicity is defined by the General Terminology Criteria for Adverse Events (CTCAE), Version 3.0, observed 5 weeks after the first cycle. If three or more toxicity grades occurred in one or more of the three patients at a given dose level, three more patients were added at that dose level. Dose limiting toxicity (DLT) was defined as the occurrence of at least three toxicity grades in at least two of six patients at that level, where the maximum tolerated dose (MTD) was determined as the dose level below one of the DLTs. Bone marrow suppression due to disease or prior treatment was evaluated, but not included in the MTD assessment.

Dose adjustments based on age and corresponding CSF volume, normal practice with therapeutic drugs administered intradural, were performed as follows: Patients under 12 months of age received a 50% dose reduction; 13- to 36-month-old patients received a 33% dose reduction; Patients over 36 months of age received full dose.

Radiologic Assessment of CNS Disease

It was not required that patients have measurable disease at study entry. Most patients have recently completed salvage radiation therapy, surgery, and / or chemotherapy, with no measurable radiological disease. Radiologic images were reviewed by a committee-certified MSK diagnostic neuro-radiologist. Since no recent evaluation of the response to neuro-oncology brain metastasis (RANO-BM) initiative was conducted at the start of the study, the assessment of radiological improvement was the first evidence of radiological improvement from the initial ¹³¹ I-8H9 treatment ²³ (measurable parenchymal disease size Reduction or improvement in augmentation for patients with meningoid disease). Swimmer graphs were generated for patients with measurable disease to indicate time to week of initial evidence of radiological improvement, length of involvement in the patient's study or follow-up, and survival. The duration of the response was assessed by calculating the time from the initial ¹³¹ I-8H9 treatment to the final radiological evaluation of the CNS disease.

Statistical evaluation

Patients were treated on an expansion cohort (25) based on the hypothesis that the introduction of ¹³¹ I-8H9 cRIT resulted in improved survival (25). Survival data was calculated from the diagnosis of CNS recurrence. Kaplan-Meier graphs were generated to assess overall survival, median survival and 95% confidence intervals (SAS, Cary, NC). Kaplan-Meier graphs were generated on subgroups of interest to assess the effect of certain factors on overall survival, including histological diagnosis, age, number of prior relapses, and disease status at study entry. Mantel-Cox analysis was performed to assess the prognostic significance of the number of injections received, total dose delivered mCi ¹³¹ I-8H9 and total dose of ¹³¹ I-8H9 delivered to CSF.

Pediatric Patients with Treatment-History with CNS Neuroblastoma Metastasis

Nineteen patients with CNS neuroblastoma were treated in MSK prior to initiation of the ¹³¹ I-8H9 cRIT treatment protocol. Survival reports in these groups and literature were used as controls in survival analysis for neuroblastoma patients treated with ¹³¹ I-8H9.

result

¹³¹ Demographics and Administration of Patients Treated with I-8H9 cRIT

134 patients were treated with 412 injections of cRIT ¹²⁴ I-8H9 and ¹³¹ I-8H9. The median age at initial cRIT injection was 8.5 years (range 1.2 to 53.5 years). Diagnosis includes metastatic CNS neuroblastoma (n = 93), sarcoma (n = 6), melanoma (n-4), ovarian carcinoma (n = 1), and primary recurrent CNS malignancy, such as medulloblastoma / PNET (n = 15). ), Ventricular meningoma (n = 9), embryonic tumors with multiple rosettes (n = 2), rod tumors (n = 1), chordomas (n = 1) and choroid plexus carcinomas (n = 2) Table 1). Two patients were excluded prior to treatment dosing due to progressive disease (neuroblastoma, N = 1) and non-compliance (ventricular tumor, N = 1). 37 patients on 10-80 mCi ¹³¹ I-8H9 (10 mCi n = 7, 20 mCi n = 3, 30 mCi n = 6, 40 mCi n = 3, 50 mCi n = 3, 60 mCi n = 4, 70 mCi n = 6, 80 mCi n = 5). The remaining 95 patients were treated with flat dose levels of an expanded cohort of 50 mCi ¹³¹ I-8H9.

Adverse Event Profile

Patients were routinely treated while awake in bed, under the direct assistance of pediatric, nuclear medicine and radiation safety teams in an outpatient setting. A rare grade 1 or 2 transient headache, fever, vomiting (acetaminophen, self-limiting with anti-emetic), manageable. The most common adverse events were prior cerebral spinal cord radiation, poor bone marrow conservation (grade 3 or 4 myelosuppression in patients with platelets <100K at entry into the study (Table 2) and in patients who received ¹³¹ I-8H9 above 50 mCi). Bone marrow suppression was expected in this massive pre-treated patient population and was noted as such, but excluded from dose limiting toxicity: Of 132 patients, 73 (59%) had 4 injections (2) as planned Received Dosimetry and 2 Total Therapies Dose) Progressive disease (n = 29), prolonged myelosuppression (n = 20), acute chemical meningitis (n = 3), subdural collection with altered CSF flow (n = 2), myelodysplasia (n = 2) and self-exclusion (n = 1) did not receive cycle # 2, which is manifested by acute headache, fever, vomiting and sterile CSF cytomegaly, Chemical meningitis was not observed in any of the patients in cycle # 1. Or it appeared in three patients after the third injection retreatment; all parties are treated with secondary care - was limited cases, it was resolved over a number of days non-myeloid inhibitory DLT is not reached (Table 2).

Dosimetric analysis

89 patients underwent CSF sampling and dosimetry as measured by ¹³¹ I-8H9 SPECT; Forty-five patients underwent dosimetry as measured by ¹²⁴ I-8H9 PET. Changes in isotope and nuclear scans were based on isotope availability and budgetary constraints, with ¹²⁴ I-8H9 PET even more expensive. For both methodologies the preferred treatment window was observed by CSF and blood sampling. CSF sampling observed inter-patient variability in total absorbed dose to CSF; The mean CSF uptake dose for the whole cohort was 104.9 cGy / mCi compared to 2.6 cGy / mCi in blood. The mean total absorbed CSF dose was 3368.8 cGy (677-13143 cGy range) by CSF sampling. Mean CSF removal was 6.7 hours. The average total therapeutic dose of ¹³¹ I-8H9 received in patients with neuroblastoma was 67.2 mCi (19.6–104.9 mCi). 65 patients evaluated for the total CSF dose delivered, including 24 who received only one treatment dose, received a total CSF dose in excess of 2100 cGy. In general, the image determination region of interest had the highest quality after ¹²⁴ I-8H9 PET compared to ¹³¹ I-8H9 SPECT and showed less inter-patient variability when compared to CSF sampling.

Neuroblastoma Subgroup Analysis

93 patients with metastatic CNS neuroblastoma received 188 tracers and treatment injections (16 dose escalation cancers; 77 expanded cohorts); 46 patients (50%) received a single treatment injection; 47 patients (50%) received two treatment injections. Upon recurrent CNS neuroblastoma, the patient underwent radiotherapy and chemotherapy, if appropriate, after biopsy or weight loss surgery. Kaplan-Meier graphs of overall survival for patients treated with ¹³¹ I-8H9 compared to pre-MSK cRIT-pre-CNS neuroblastoma patients are provided in FIG. 1. Patients were analyzed in two groups: Group 1 patients included systemic immunotherapy using intravenous MoAb 3F8 and GMCSF as described above in addition to radiotherapy, temozolamide / irinotecan, cRIT ¹³¹ I-8H9 (13). Have undergone full CNS and systemic induction therapy. Group 2 patients were treated with all other therapies and cRIT ¹³¹ I-8H9 (FIG. 2).

^Of the 93 patients treated with ¹³¹ I-8H9 cRIT, 42 (53%) remained alive from the time of CNS metastasis until the date of data cutoff (range: 4.8-152.4 months). Of the 93 patients, 98% survived at least 6 months, 88% survived at least 12 months, 71% survived at least 36 months, and 51% survived more than 60 months. Median survival for patients treated with ¹³¹ I-8H9 cRIT was 31.8 (3.8-170.1) months (95% confidence interval [CI] lower limit: 35.2 months). In contrast, of 19 MSK cRIT-pre patients, 6 (32%) survived at least 6 months, 4 (21%) survived at least 12 months, and 2 (11%) survived more than 36 months. However, no one survived to 60 months. Overall survival of the MSK control cohort ranged from 2 days to 44.1 months and had an estimated median survival of 5.5 months (95% CI: 1.1 to 8.7 months).

Eighteen of the 93 patients (47%) died of disease-associated CNS recurrence—CNS relapse alone (11 patients) or both CNS and systemic relapse (7 patients). Sixteen patients (42%) died of recurrence of systemic disease. Four patients (11%) died from causes other than neuroblastoma. The proportion of patients in whom death was not associated with recurrence of CNS disease (20/38, 53%) provides further evidence of the effectiveness of ¹³¹ I-8H9 therapy in the treatment of CNS neuroblastoma. These patients survived up to 89.8 months after their first ¹³¹ I-8H9 treatment without recurring its CNS disease. After initiating ¹³¹ I-8H9 therapy, 11 survived for more than 1 year and 4 survived for at least 2 years.

Results for Infants with CNS Neuroblastoma

CNS metastasis occurred in 18 patients with neuroblastoma initially diagnosed under 18 months of age; Twelve (66%) had MYCN amplified tumors. Patients developed CNS neuroblastoma as a site of disease recurrence (N = 13) or in an intractable systemic neuroblastoma environment (N = 5). Three patients developed symptomatic neuroblastoma and advanced and died of systemic (N = 2) or CNS (n = 1) neuroblastoma before initiation of cRIT131I-8H9. Of the remaining 15, 12 received a CNS rescue plan that included radiotherapy, chemotherapy and cRIT131I-8H9. Overall survival after cRIT 131I-8H9 was significantly prolonged in the young patient subset compared to patients initially diagnosed above 18 months of age (p = 0.0096;) (FIG. 3). Twelve patients, including one patient with hundreds of neuroblastoma metastases across the dural space (FIG. 5), remain on average 6.3 years (1.6-11.8 years) as long-term survivors from detection of CNS disease (FIG. 3).

Based on Radiologic Assessment in Patients with CNS Neuroblastoma ¹³¹ Validity of I-8H9 cRIT

^Of the 93 patients treated with ¹³¹ I-8H9 cRIT, 21 (23%) had radiological evidence of CNS / LM disease at the time they received their first ¹³¹ I-8H9 treatment. 4A-4B provide a table summary of the radiological evaluation results after initial treatment for these patients. Of these, 9 (43%) showed a radiological improvement (reduced lesion size and / or reduced meningeal augmentation) and 9 patients (43%) showed stable disease upon initial radiological evaluation. Additional objective evidence of the long-term clinical benefit of ¹³¹ I-8H9 therapy may be provided by the sustained analysis of radiological stability. ^While median survival (58 months) for 93 patients treated with ¹³¹ I-8H9 significantly exceeded median survival (5.5 months) for past patients, a significant improvement in survival was necessarily associated with long-term, persistent sedation of the disease. It may not be. Treatment with ¹³¹ I-8H9 induced a sustained response in a large number of patients. Based on the available data, the median duration of radiological response is 49 weeks (95% CI: 32.1-73.7 weeks), ranging from 2.6 weeks to 676 weeks (13 years). The median duration of radiological response is likewise considerably downgraded, but several patients survived much longer than their last scan date. Nevertheless, the median duration (11.3 months) of the radiological response to ¹³¹ I-8H9 treatment exceeds the past median survival of 5.5 months (FIGS. 4A-4B).

Subgroup Analysis and Effects on Overall Survival

Several potentially potentially life-threats that might not have been captured in the overall survival analysis, including known demographic and genetic risk factors, disease characteristics at study entry, the number of relapses before ¹³¹ I-8H9, and additional therapies received after cRIT 8H9. Factors exist To determine the effect of these risk factors on the overall survival of patients treated with ¹³¹ I-8H9, subgroup analysis was performed.

In the neuroblastoma cohort, several risk factors, including age at diagnosis, INSS stage, and MYCN amplification status, were consistently associated with progression and overall survival of neuroblastoma. Age at initial diagnosis was prognostic, but overall survival of patients with MYCN amplified tumors was not (p = 0.2490). To determine if there is any difference in patient survival based on the time points at which treatment was received (ie, the first half of the study period [2004-2009] versus the second half [2010-present]. There was no difference in survival of treated CNS neuroblastoma patients during the initial period compared to treated CNS neuroblastoma patients (p = 0.8851) ¹³¹ I-8H9 cRIT pre-CSI had no effect on overall survival in these patients. There was also no statistical difference when 21 Gy CSI vs 18 Gy CSI was administered (Figure 6), CSF sampling was given more than 50 mCi ¹³¹ I-8H9 or greater than 2100 cGy. It was also noted that there was no statistical difference in survival among recipients, although not statistically significant (p = 0.08), but there was a trend toward improved survival in patients receiving two ¹³¹ I-8H9 treatment injections. This was noted.

In another patient cohort, in 6/15 of patients with recurrent medulloblastoma, 3/9 of patients with recurrent ventricular meningoma, 2 patients with embryonic tumors with multilayer rosettes and in 1 patient with recurrent choroidal plexus carcinoma Prolonged survival was observed (Table 4).

Overall, 134 patients (93 neuroblastomas, 11 other non-CNS tumors and 30 primary CNS tumors) received 412 injections. The mean absorbed dose was 104.9 cGy / mCi in CSF and 2.6 cGy / mCi in blood. Acute toxicity was limited. Although not dose-limiting toxicity, grade 3 or 4 myelosuppression has been noted in patients with prior cerebral spinal cord radiation therapy and doses of 60 mCi ¹³¹ I-8H9 or higher. Improved OS has been noted for neuroblastoma cohorts as reported by conventional therapies.

Argument

Targeting of checkpoint modulators of the B7 family is at the forefront of cancer research and has strong evidence demonstrating the major regulatory role of B7-H3 on T-cell proliferation. B7-H3 expression is significantly associated with poor outcomes in some cancers and is uniquely overexpressed in cancer as compared to normal human tissue. Maturation data (up to 13 years) of well-tolerated cRIT-based therapy including compartmental radiolabeled anti-B7-H3 for incurable embryo tumors in the pediatric population was presented. Preferred adverse event profiles, therapeutic indices, and prolonged survival for some histological diagnoses, including treated recurrent CNS neuroblastoma, medulloblastoma, ventricular tumor, choroid plexus carcinoma, and embryonic tumors with multilayer rosettes, are encouraging. CRITs using ¹²⁴ I-8H9 and ¹³¹ I-8H9 in the ventricle are very young and appear to be manageable, acutely toxic and safe, even in large pre-treated patient populations. The basis of the secondary mechanism for successful eradication of microscopic tumor cells may be due in part to 8H9 complement activation into the CSF space, but tumor cell cytotoxicity is due to the direct effect of ¹³¹ I-radiation. Complement components C3 and C5b-9 were found to be present in CSF following intraventricular rituximab for recurrent CNS lymphoma (17). More recent evidence suggests that a widespread known tumor vasculature overexpresses B7-H3; Targeting the tumor vasculature may have additional therapeutic benefits.

In the past and in the present, the prognosis for pediatric patients diagnosed with recurrent CNS neuroblastoma remains poor, with currently available treatment options. Expected survival reported across sites and countries typically does not exceed 6 months, and long-term survival beyond 3 years occurs in less than 10% of patients (18-20). Past patient cohorts from the current study were consistent with this previous study, which reported a short survival time and poor prognosis after CNS metastasis. The median overall survival of patients treated with MSK prior to the onset of cRIT in 2004 was 5.5 months; Only 2 of 19 patients survived for at least 36 months, and none survived beyond 44 months. Regardless of the patient's treatment or geography, these figures have not changed significantly over the last 40 years. Introduction of cRIT ¹³¹ I-8H9 represents a significant advance in the treatment of this disease and has significant improvement in median overall survival and radical.

Since the number of CNS neuroblastoma survivors has increased over the years, the goal was to minimize the risks associated with known cerebrospinal radiation, most importantly neurocognitive deficiency, endocrine disease and growth retardation in young children. The data suggest that the degree of neurocognitive impairment (ie, mild, severity, or severe) demonstrates a dose-response pattern (11). The trend further suggests that the combination of cerebrospinal radiation and cRIT can eradicate bulky meningeal deposition that is not suitable for surgical resection. Half of the patients in the cRIT- rescue therapy cohort were treated with 2160 cGy CSI, the standard dose administered for national control of neuroblastoma primary tumors. Since the target cGY delivered by cRIT has increased since 2009, the CSI dose representing half of the patients in this cohort has been reduced to 1800 cGy. The data did not demonstrate a recognizable reduction in the effects of treatment with combination external beam radiation therapy 1800 cGy and cRIT. In addition, because of prior radiotherapy, very young age, or parental choices at the time of initial diagnosis, five patients received CSI below 18 Gy, but maintained long-term CNS disease control. Although only pursued more recently, proton-beam radiotherapy delivers significantly less radiation to healthy tissues than conventional proton therapy, and may be an additional way to minimize long-term morbidity. Reduced-dose cerebrospinal irradiation and cRIT targeting micrometastatic neuroblastomas aimed at controlling bulky parenchymal and nodular meningoid disease may be sufficient to prolong survival.

A further focus of the study is to determine the optimal cGy delivered to CSF by cRIT ¹³¹ I-8H9 to completely eradicate micrometastatic deposition. Since the patient initially enrolled in phase I studies with ¹³¹ I-8H9, the dose of 8H9 changed. The patients were then all enrolled in a phase II cohort expanded with a uniform treatment dose of 50 mCi ¹³¹ I-8H9. The mean dose to the bone marrow was clearly consistent in all patients, but the cGy / mCi dose delivered by cRIT to CSF was very variable. This may be a reflection of the variable rate of CSF flow based on the presence or absence of pre-surgical, radiation, bulky lesions, all leading to inherent CSF flow differences before cRIT. Since most patients have been treated with cRIT as a complication for micrometastasis, the fraction of injected antibodies needed to eradicate tumor lesions is also difficult to assess. In addition, other immuno-PET studies in patients with brain metastasis suggest that antibody uptake may be highly variable in different lesions of the same patient (12). No differences in survival were identified by CSF sampling for patients awarded ¹³¹ I-8H9> 50 mCi with CSF, or for patients awarded> 2100 cGy. This suggests that higher doses of cRIT may not be needed unless lower simultaneous CSI doses are being considered.

Attempts have been made to determine which patients are at risk of developing recurrent CNS disease. Risk factors include diagnostic lumbar puncture and MYCN amplification at initial neuroblastoma diagnosis (19, 20). The identification of genomic mutations that induce the metastasis process has been a challenge. In SNP analysis of CNS metastasis and its pre-CNS-primary tumor pairs, only a few specific and recurrent differences in genomic lesions were identified (21). However, in a series of 13 CNS neuroblastoma metastases with corresponding primary tumors, we have previously shown that miR-29a may be a biomarker for CNS metastasis; Downregulation may play a pivotal role in CNS progression (22). Known tumor targets of miR-29a included CDC6, CDK6 and DNMT3A and B7-H3. These targets were found to have higher differential expression in brain metastases than their paired primary (22). Prophylactic treatments using well-tolerated cRIT based therapies for patients at high risk for recurrence should be explored.

CRIT to ¹³¹ I-8H9 is safe, has desirable dosimetry in CSF and blood, and is promising for improving the prognosis of malignancies involving the CNS. Metastatic tumors on the CNS can be completely eradicated, removing the site of protection against malignancies. Intraventricular radioimmunotherapy can be successfully included in curative treatment strategies for some pediatric histology, including CNS neuroblastoma. These data support the role for other high-risk tumors, including recurrent medulloblastoma, ventricular melanoma, choroid plexus carcinoma, embryonic tumors with multilayer rosettes. Overall, 60% of patients with CNS neuroblastoma metastases achieve long-term sedation, including 33% of patients with multiple parenchymal metastases. Survival benefits are shown for patients with B7-H3 overexpressing tumors treated with CNS induction therapy with cRIT ¹³¹ I-8H9.

Histological diagnosis Diagnosis Number of patients Number of pts on one phase (10 to 80 mCi) Pts number of extended cohort (50 mCi) Injection count Neuroblastoma 93 * 16 77 293 Medulloblastoma / PNET 15 6 9 29 Ventricular ductoma 9+ 4 5 37 ETMR 2 One One 4 sarcoma 6 3 3 18 Melanoma 4 3 One 9 Other (ATRT, choriofreezing carcinoma, ovarian carcinoma, retinoblastoma) 5 3 2 22 gun 134 ** 36 98 412

* One patient was removed due to progressive disease before receiving the treatment injection

+ 1 patient removed due to non-compliance before receiving a therapeutic injection

** Therapies are injected into 132 people

Adverse Event Profile by Histology Diagnosis Number Possible or probable adverse events (CTC 3.0) Bone marrow suppression (%) Neuroblastoma 93 Grade 3 or 4 myelosuppression (ANC, hgb, platelets) (83)
Grade 4 hypersensitivity reactions (1)
Class 3 ALT / AST (5)
Grade 3 Chemical Meningitis (3)
Grade 4 MDS / AML (5) 83 (89%) Medulloblastoma /
PNET 15 Grade 3 or 4 myelosuppression (6)
Grade 4 Chemical Meningitis (1) 6 (43%) Ventricular ductoma 9 Grade 3 or 4 myelosuppression (3) 3 (33%) ETMR
2 Grade 3 or 4 myelosuppression (2) 2 (100%) sarcoma 6 Grade 3 or 4 myelosuppression (3)
Grade 4 AML (1) 3 (50%) Melanoma 4 Grade 3 bone marrow suppression (2)
Level 3 Coming (1)
Grade 3 hypokalemia (1) 2 (50%) Other (ATRT, choriofreezing carcinoma, ovarian carcinoma, retinoblastoma) 5 Grade 4 MDS / AML (1) gun 132

Characteristic neuroblastoma Cohort N = 93 Age, age at first diagnosis Median (three, range) 2.98 (1 day to 12 years old) Age, age at first cRIT injection Median (three, range) 4.65 (16 months to 13 years old) Steps at first diagnosis 4 90 3, 4s 3 MYCN-amplified neuroblastoma 46 (49%) Cerebrospinal radiation therapy for CNS diagnosis > 2160 cGY 3 (3%) 2160 cGy 30 (32%) 1800 cGy 44 (47%) <18 cGy 7 (8%) Topical only or no CSI 9 (10%) Total CNS Radiation, Chemotherapy, and cRIT-8H9 (Group 1) 64 (69%) cRIT-8H9 and all other therapies (group 2) 29 (31%) cRIT 8H9 ex relapse # 0 55 (59%) One 18 (19%) 2 1 (1%) 3 0 4 1 (1%) Refractory body 17 (18%) CNS Disease Types Monotopic substance 54 (58%) Multilocal Substance 21 (23%) Water curtain 9 (10%) Real + water film 9 (10%) Symptomatic CNS Neuroblastoma 59 (63%) Conditions when treated with cRIT 8H9: Stable evaluable disease 21 (23%) Radiologic / Cytologic Sedation 72 (77%)

Numbers are indicated in percent in parentheses unless otherwise noted

Other embryos and other CNS malignancies For survival Diagnosis Number of patients Overall survival (month) Medulloblastoma 15 8.2 (1-100) Ventricular ductoma 9 13.3 (2.8-117) ETMR 2 34 (2053) sarcoma 6 10 (0.7-46) Melanoma 4 5 (0.6 ~ 7.3) Other (ATRT, CPP, Ovarian Carcinoma, RB, Chordoma) 5 9.3 to 98

references

1. Wiens AL, Hattab EM. The pathologic spectrum of solid CNS metastases in the pediatric population. J Neurosurg Pediatr 2014, 14 (2): 129-135.

2. De B, Kinnaman MD, Wexler LH, Kramer K, Wolden SL. Central nervous system relapse of rhabdomyosarcoma. Pediatr Blood Cancer. 2018 Jan; 65 (1). doi: 10.1002 / pbc.26710. Epub 2017 Jul 11.

Pizzo ME, Wolak DJ, Kumar NN, Brunette E, Brunnquell CL, Hannocks MJ, Abbott NJ, Meyerand ME, Sorokin L, Stanimirovic DB, Thorne RG. Intrathecal antibody distribution in the rat brain: surface diffusion, perivascular transport and osmotic enhancement of delivery. J Physiol. 2018 Feb 1; 596 (3): 445-475. doi: 10.1113 / JP275105. Epub 2017 Dec 18.

4.Convection-Enhanced Delivery for Diffuse Intrinsic Pontine Glioma Treatment. Zhou Z, Singh R, Souweidane MM. Curr Neuropharmacol. 2017; 15 (1): 116-128.

Kramer K, Smith-Jones PM, Humm JL, Zanzonico P, Pandit-Taskar N, Carrasquillo J, Modak S, Tickoo S, Gerald WL, Dunkel IJ, Khakoo Y, Gilheeney SW, Souweidane MM, Larson SM, and Cheung NKV. Radioimmunotherapy for High-Risk and Recurrent Central Nervous System (CNS) Cancers: Results of a Phase II Study with intra-Ommaya ¹³¹ I-3F8. NeuroOncology 2010; 12 (6): ii48.

Kramer K, Kushner BH, Modak S, Pandit-Taskar N, Smith-Jones P, Zanzonico P, Humm JL, Xu H, Wolden SL, Souweidane MM, Larson SM, Cheung NK. Compartmental intrathecal radioimmunotherapy: results for treatment for metastatic CNS neuroblastoma. J Neurooncol 2010; 97 (3): 409-187.

7. Mehta AI, Choi BD, Ajay D, Raghavan R, Brady M, Friedman AH, Pastan I, Bigner DD, Sampson JH. Convection enhanced delivery of macromolecules for brain tumors. Curr Drug Discov Technol. 2012 Dec; 9 (4): 305-10.

8. Pizer BL, Papanastassiou V, Moseley R, Tzanis S, Hancock JP, Kemshead JT, et al. Meningeal Leukemia and Medulloblastoma-Preliminary Experience with Intrathecal Radioimmunotherapy. Antibody Immunoconj. 1991; 4 (4): 753-61.

9. Bigner DD, Brown MT, Friedman AH, Coleman RE, Akabani G, Friedman HS, et al. Iodine-131-labeled antitenascin monoclonal antibody 81C6 treatment of patients with recurrent malignant gliomas: phase I study results. Journal of clinical oncology: official journal of the American Society of Clinical Oncology. 1998; 16 (6): 2202-12.

10. Cokgor I, Akabani G, Friedman HS, Friedman AH, Zalutsky MR, Zehngebot LM, et al. Long term response in a patient with neoplastic meningitis secondary to melanoma treated with (131) I-radiolabeled antichondroitin proteoglycan sulfate Mel-14 F (ab ') (2): a case study. Cancer. 2001; 91 (9): 1809-13.

11. Kramer K, Humm JL, Souweidane MM, Zanzonico PB, Dunkel IJ, Gerald WL, et al. Phase I study of targeted radioimmunotherapy for leptomeningeal cancers using intra-Ommaya 131-I-3F8. Journal of clinical oncology: official journal of the American Society of Clinical Oncology. 2007; 25 (34): 5465-70.

12. Modak S, Kramer K, Gultekin SH, Guo HF, Cheung NK. Monoclonal antibody 8H9 targets a novel cell surface antigen expressed by a wide spectrum of human solid tumors. Cancer Res. 2001; 61 (10): 4048-54.

13.Modak S, Guo HF, Humm JL, Smith-Jones PM, Larson SM, Cheung NK. Radioimmunotargeting of human rhabdomyosarcoma using monoclonal antibody 8H9. Cancer Biother Radiopharm. 2005; 20 (5): 534-46.

14. Miraldi FD, Nelson AD, Kraly C, Ellery S, Landmeier B, Coccia PF, et al. Diagnostic imaging of human neuroblastoma with radiolabeled antibody. Radiology. 1986; 161 (2): 413-8.

15. Brodeur GM, Seeger RC, Barrett A, Berthold F, Castleberry RP, D'Angio G, et al. International criteria for diagnosis, staging, and response to treatment in patients with neuroblastoma. J Clin Oncol. 1988; 6 (12): 1874-81.

16. Monclair T, Brodeur GM, Ambros PF, Brisse HJ, Cecchetto G, Holmes K, et al. The International Neuroblastoma Risk Group (INRG) Staging System: An INRG Task Force Report. Journal of Clinical Oncology. 2009; 27 (2): 298-303.

17. Kadoch C, Li J, Wong VS, Chen L, Cha S, Munster P, et al. Complement activation and intraventricular rituximab distribution in recurrent central nervous system lymphoma. Clinical cancer research: an official journal of the American Association for Cancer Research. 2014; 20 (4): 1029-41.

18. Kellie SJ, Hayes FA, Bowman L, Kovnar EH, Langston J, Jenkins JJ, 3rd, et al. Primary extracranial neuroblastoma with central nervous system metastases characterization by clinicopathologic findings and neuroimaging. Cancer. 1991; 68 (9): 1999-2006.

19. Kramer K, Kushner B, Heller G, Cheung NK. Neuroblastoma metastatic to the central nervous system. The Memorial Sloan-kettering Cancer Center Experience and A Literature Review. Cancer. 2001; 91 (8): 1510-9.

20. Matthay KK, Brisse H, Couanet D, Couturier J, Benard J, Mosseri V, et al. Central nervous system metastases in neuroblastoma: radiographic, clinical, and biologic features in 23 patients. Cancer. 2003; 98 (1): 155-65.

21. Cobrinik D, Ostrovnaya I, Hassimi M, Tickoo SK, Cheung IY, Cheung NK. Recurrent pre-existing and acquired DNA copy number alterations, including focal TERT gains, in neuroblastoma central nervous system metastases. Genes, chromosomes & cancer. 2013; 52 (12): 1150-66.

22. Cheung IY, Farazi TA, Ostrovnaya I, Xu H, Tran H, Mihailovic A, et al. Deep MicroRNA sequencing reveals downregulation of miR-29a in neuroblastoma central nervous system metastasis. Genes, chromosomes & cancer. 2014; 53 (10): 803-14.

Alexander BM, Brown PD, Ahluwalia MS, Aoyama H, Baumert BG, Chang SM, Gaspar LE, Kalkanis SN, Macdonald DR, Mehta MP, Soffietti R, Suh JH, van den Bent MJ, Vogelbaum MA, Wefel JS, Lee EQ, Wen PY; Response Assessment in Neuro-Oncology (RANO) group. Clinical trial design for local therapies for brain metastases: a guideline by the Response Assessment in Neuro-Oncology Brain Metastases working group. Lancet Oncol. 2018 Jan; 19 (1): e33-e42. doi: 10.1016 / S1470-2045 (17) 30692-7.

Kramer K, Pandit-Taskar N, Humm JL, Zanzonico PB, Haque S, Dunkel IJ, Wolden SL, Donzelli M, Goldman DA, Lewis JS, Lyashchenko SK, Khakoo Y, Carrasquillo JA, Souweidane MM, Greenfield JP, Lyden D, De Braganca KD, Gilheeney SW, Larson SM, Cheung NKV. A phase II study of radioimmunotherapy with intraventricular 131I-3F8 for medulloblastoma. Pediatr Blood Cancer. 2017 Sep 22.doi: 10.1002 / pbc.26754. (PMID: 28940863)

25. Kramer K, Kushner BH, Modak S, Pandit-Taskar N, Smith-Jones P, Zanzonico P, Humm JL, Xu H, Wolden SL, Souweidane MM, Larson SM, Cheung NK. Compartmental intrathecal radioimmunotherapy: results for treatment for metastatic CNS neuroblastoma. Journal of Neurooncology, 97: 409-418, 2010. (PMID: 19890606)

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Tyr   1 5 10 <210> 5 <211> 9 <212> PRT <213> Mus musculus <400> 5 Gln Thr Thr Ala Thr Trp Phe Ala Tyr   1 5 <210> 6 <211> 11 <212> PRT <213> Mus musculus <400> 6 Arg Ala Ser Gln Ser Ile Ser Asp Tyr Leu His   1 5 10 <210> 7 <211> 7 <212> PRT <213> Mus musculus <400> 7 Tyr Ala Ser Gln Ser Ile Ser   1 5 <210> 8 <211> 9 <212> PRT <213> Mus musculus <400> 8 Gln Asn Gly His Ser Phe Pro Leu Thr   1 5 <210> 9 <211> 243 <212> PRT <213> Mus musculus <400> 9 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala   1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr              20 25 30 Asp Ile Asn Trp Val Arg Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile          35 40 45 Gly Trp Ile Phe Pro Gly Asp Gly Ser Thr Gln Tyr Asn Glu Lys Phe      50 55 60 Lys Gly Lys Ala Thr Leu Thr Thr Asp Thr Ser Ser Ser Thr Ala Tyr  65 70 75 80 Met Gln Leu Ser Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys                  85 90 95 Ala Arg Gln Thr Thr Ala Thr Trp Phe Ala Tyr Trp Gly Gln Gly Thr             100 105 110 Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser         115 120 125 Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Thr Thr Leu     130 135 140 Ser Val Thr Pro Gly Asp Arg Val Ser Leu Ser Cys Arg Ala Ser Gln 145 150 155 160 Ser Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser                 165 170 175 Pro Arg Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro             180 185 190 Ser Arg Phe Ser Gly Ser Gly Ser Gly Ser Asp Phe Thr Leu Ser Ile         195 200 205 Asn Ser Val Glu Pro Glu Asp Val Gly Val Tyr Tyr Cys Gln Asn Gly     210 215 220 His Ser Phe Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 225 230 235 240 Gln ala ala             <210> 10 <211> 731 <212> DNA <213> Mus musculus <400> 10 caggtcaaac tgcagcagtc tggggctgaa ctggtaaagc ctggggcttc agtgaaattg 60 tcctgcaagg cttctggcta caccttcaca aactatgata taaactgggt gaggcagagg 120 cctgaacagg gacttgagtg gattggatgg atttttcctg gagatggtag tactcaatac 180 aatgagaagt tcaagggcaa ggccacactg actacagaca catcctccag cacagcctac 240 atgcagctca gcaggctgac atctgaggac tctgctgtct atttctgtgc aagacagact 300 acggctacct ggtttgctta ctggggccaa gggaccacgg tcaccgtctc ctcaggtgga 360 ggcggttcag gcggaggtgg ctctggcggt ggcggatcgg acatcgagct cactcagtct 420 ccaaccaccc tgtctgtgac tccaggagat agagtctctc tttcctgcag ggccagccag 480 agtattagcg actacttaca ctggtaccaa caaaaatcac atgagtctcc aaggcttctc 540 atcaaatatg cttcccaatc catctctggg atcccctcca ggttcagtgg cagtggatca 600 gggtcagatt tcactctcag tatcaacagt gtggaacctg aagatgttgg agtgtattac 660 tgtcaaaatg gtcacagctt tccgctcacg ttcggtgctg ggaccaagct ggagctgaaa 720 caggcggccg c 731 <210> 11 <211> 731 <212> DNA <213> Mus musculus <400> 11 gtccagtttg acgtcgtcag accccgactt gaccatttcg gaccccgaag tcactttaac 60 aggacgttcc gaagaccgat gtggaagtgt ttgatactat atttgaccca ctccgtctcc 120 ggacttgtcc ctgaactcac ctaacctacc taaaaaggac ctctaccatc atgagttatg 180 ttactcttca agttcccgtt ccggtgtgac tgatgtctgt gtaggaggtc gtgtcggatg 240 tacgtcgagt cgtccgactg tagactcctg agacgacaga taaagacacg ttctgtctga 300 tgccgatgga ccaaacgaat gaccccggtt ccctggtgcc agtggcagag gagtccacct 360 ccgccaagtc cgcctccacc gagaccgcca ccgcctagcc tgtagctcga gtgagtcaga 420 ggttggtggg acagacactg aggtcctcta tctcagagag aaaggacgtc ccggtcggtc 480 tcataatcgc tgatgaatgt gaccatggtt gtttttagtg tactcagagg ttccgaagag 540 tagtttatac gaagggttag gtagagaccc taggggaggt ccaagtcacc gtcacctagt 600 cccagtctaa agtgagagtc atagttgtca caccttggac ttctacaacc tcacataatg 660 acagttttac cagtgtcgaa aggcgagtgc aagccacgac cctggttcga cctcgacttt 720 gtccgccggc g 731 <210> 12 <211> 731 <212> DNA <213> Mus musculus <400> 12 caggtcaaac tgcagcagtc tggggctgaa ctggtaaagc ctggggcttc agtgaaattg 60 tcctgcaagg cttctggcta caccttcaca aactatgata taaactgggt gaggcagagg 120 cctgaacagg gacttgagtg gattggatgg atttttcctg gagatggtag tactcaatac 180 aatgagaagt tcaagggcaa ggccacactg actacagaca catcctccag cacagcctac 240 atgcagctca gcaggctgac atctgaggac tctgctgtct atttctgtgc aagacagact 300 acggctacct ggtttgctta ctggggccaa gggaccacgg tcaccgtctc ctcagatgga 360 ggcggttcag gcggaggtgg ctctggcggt ggcggatcgg acatcgagct cactcagtct 420 ccaaccaccc tgtctgtgac tccaggagat agagtctctc tttcctgcag ggccagccag 480 agtattagcg actacttaca ctggtaccaa caaaaatcac atgagtctcc aaggcttctc 540 atcaaatatg cttcccaatc catctctggg atcccctcca ggttcagtgg cagtggatca 600 gggtcagatt tcactctcag tatcaacagt gtggaacctg aagatgttgg agtgtattac 660 tgtcaaaatg gtcacagctt tccgctcacg ttcggtgctg ggaccaagct ggagctgaaa 720 caggcggccg c 731 <210> 13 <211> 243 <212> PRT <213> Mus musculus <400> 13 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala   1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr              20 25 30 Asp Ile Asn Trp Val Arg Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile          35 40 45 Gly Trp Ile Phe Pro Gly Asp Gly Ser Thr Gln Tyr Asn Glu Lys Phe      50 55 60 Lys Gly Lys Ala Thr Leu Thr Thr Asp Thr Ser Ser Ser Thr Ala Tyr  65 70 75 80 Met Gln Leu Ser Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys                  85 90 95 Ala Arg Gln Thr Thr Ala Thr Trp Phe Ala Tyr Trp Gly Gln Gly Thr             100 105 110 Thr Val Thr Val Ser Ser Asp Gly Gly Gly Ser Gly Gly Gly Gly Ser         115 120 125 Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Thr Thr Leu     130 135 140 Ser Val Thr Pro Gly Asp Arg Val Ser Leu Ser Cys Arg Ala Ser Gln 145 150 155 160 Ser Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser                 165 170 175 Pro Arg Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro             180 185 190 Ser Arg Phe Ser Gly Ser Gly Ser Gly Ser Asp Phe Thr Leu Ser Ile         195 200 205 Asn Ser Val Glu Pro Glu Asp Val Gly Val Tyr Tyr Cys Gln Asn Gly     210 215 220 His Ser Phe Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 225 230 235 240 Gln ala ala             <210> 14 <211> 243 <212> PRT <213> Mus musculus <400> 14 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Glu Pro Gly Ala   1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr              20 25 30 Asp Ile Asn Trp Val Arg Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile          35 40 45 Gly Trp Ile Phe Pro Gly Asp Gly Ser Thr Gln Tyr Asn Glu Lys Phe      50 55 60 Lys Gly Lys Ala Thr Leu Thr Thr Asp Thr Ser Ser Ser Thr Ala Tyr  65 70 75 80 Met Gln Leu Ser Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys                  85 90 95 Ala Arg Gln Thr Thr Ala Thr Trp Phe Ala Tyr Trp Gly Gln Gly Thr             100 105 110 Thr Val Thr Val Ser Ser Asp Gly Gly Gly Ser Gly Gly Gly Gly Ser         115 120 125 Gly Gly Gly Gly Ser Asp Ile Glu Leu Thr Gln Ser Pro Thr Thr Leu     130 135 140 Ser Val Thr Pro Gly Asp Gln Val Ser Leu Ser Cys Arg Ala Ser Gln 145 150 155 160 Ser Ile Ser Asp Tyr Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser                 165 170 175 Pro Gln Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro             180 185 190 Ser Arg Phe Ser Gly Ser Gly Ser Gly Ser Asp Phe Thr Leu Ser Ile         195 200 205 Asn Ser Val Glu Pro Glu Asp Val Gly Val Tyr Tyr Cys Gln Asn Gly     210 215 220 His Ser Phe Pro Leu Thr Phe Gly Ala Gly Thr Glu Leu Glu Leu Glu 225 230 235 240 Gln ala ala             <210> 15 <211> 18 <212> PRT <213> Mus musculus <400> 15 Asn Pro Val Leu Gln Gln Asp Ala His Ser Ser Val Thr Ile Thr Pro   1 5 10 15 Gln arg         <210> 16 <211> 26 <212> PRT <213> Mus musculus <400> 16 Ser Pro Thr Gly Ala Val Glu Val Gln Val Pro Glu Asp Pro Val Val   1 5 10 15 Ala Leu Val Gly Thr Asp Ala Thr Leu Arg              20 25 <210> 17 <211> 534 <212> PRT <213> Mus musculus <400> 17 Met Leu Arg Arg Arg Gly Ser Pro Gly Met Gly Val His Val Gly Ala   1 5 10 15 Ala Leu Gly Ala Leu Trp Phe Cys Leu Thr Gly Ala Leu Glu Val Gln              20 25 30 Val Pro Glu Asp Pro Val Val Ala Leu Val Gly Thr Asp Ala Thr Leu          35 40 45 Cys Cys Ser Phe Ser Pro Glu Pro Gly Phe Ser Leu Ala Gln Leu Asn      50 55 60 Leu Ile Trp Gln Leu Thr Asp Thr Lys Gln Leu Val His Ser Phe Ala  65 70 75 80 Glu Gly Gln Asp Gln Gly Ser Ala Tyr Ala Asn Arg Thr Ala Leu Phe                  85 90 95 Pro Asp Leu Leu Ala Gln Gly Asn Ala Ser Leu Arg Leu Gln Arg Val             100 105 110 Arg Val Ala Asp Glu Gly Ser Phe Thr Cys Phe Val Ser Ile Arg Asp         115 120 125 Phe Gly Ser Ala Ala Val Ser Leu Gln Val Ala Ala Pro Tyr Ser Lys     130 135 140 Pro Ser Met Thr Leu Glu Pro Asn Lys Asp Leu Arg Pro Gly Asp Thr 145 150 155 160 Val Thr Ile Thr Cys Ser Ser Tyr Gln Gly Tyr Pro Glu Ala Glu Val                 165 170 175 Phe Trp Gln Asp Gly Gln Gly Val Pro Leu Thr Gly Asn Val Thr Thr             180 185 190 Ser Gln Met Ala Asn Glu Gln Gly Leu Phe Asp Val His Ser Ile Leu         195 200 205 Arg Val Val Leu Gly Ala Asn Gly Thr Tyr Ser Cys Leu Val Arg Asn     210 215 220 Pro Val Leu Gln Gln Asp Ala His Ser Ser Val Thr Ile Thr Pro Gln 225 230 235 240 Arg Ser Pro Thr Gly Ala Val Glu Val Gln Val Pro Glu Asp Pro Val                 245 250 255 Val Ala Leu Val Gly Thr Asp Ala Thr Leu Arg Cys Ser Phe Ser Pro             260 265 270 Glu Pro Gly Phe Ser Leu Ala Gln Leu Asn Leu Ile Trp Gln Leu Thr         275 280 285 Asp Thr Lys Gln Leu Val His Ser Phe Thr Glu Gly Arg Asp Gln Gly     290 295 300 Ser Ala Tyr Ala Asn Arg Thr Ala Leu Phe Pro Asp Leu Leu Ala Gln 305 310 315 320 Gly Asn Ala Ser Leu Arg Leu Gln Arg Val Arg Val Ala Asp Glu Gly                 325 330 335 Ser Phe Thr Cys Phe Val Ser Ile Arg Asp Phe Gly Ser Ala Ala Val             340 345 350 Ser Leu Gln Val Ala Ala Pro Tyr Ser Lys Pro Ser Met Thr Leu Glu         355 360 365 Pro Asn Lys Asp Leu Arg Pro Gly Asp Thr Val Thr Ile Thr Cys Ser     370 375 380 Ser Tyr Arg Gly Tyr Pro Glu Ala Glu Val Phe Trp Gln Asp Gly Gln 385 390 395 400 Gly Val Pro Leu Thr Gly Asn Val Thr Thr Ser Gln Met Ala Asn Glu                 405 410 415 Gln Gly Leu Phe Asp Val His Ser Val Leu Arg Val Val Leu Gly Ala             420 425 430 Asn Gly Thr Tyr Ser Cys Leu Val Arg Asn Pro Val Leu Gln Gln Asp         435 440 445 Ala His Gly Ser Val Thr Ile Thr Gly Gln Pro Met Thr Phe Pro Pro     450 455 460 Glu Ala Leu Trp Val Thr Val Gly Leu Ser Val Cys Leu Ile Ala Leu 465 470 475 480 Leu Val Ala Leu Ala Phe Val Cys Trp Arg Lys Ile Lys Gln Ser Cys                 485 490 495 Glu Glu Glu Asn Ala Gly Ala Glu Asp Gln Asp Gly Glu Gly Glu Gly             500 505 510 Ser Lys Thr Ala Leu Gln Pro Leu Lys His Ser Asp Ser Lys Glu Asp         515 520 525 Asp Gly Gln Glu Ile Ala     530

Claims (63)

A method of treating central nervous system (CNS) cancer in a human subject, comprising administering a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds human B7H3 into the subject's CNS,
The cancer is a primary central nervous system (CNS) cancer or a cancer metastatic to the CNS, and wherein the antibody or antigen-binding fragment thereof is conjugated to a radioisotope and / or mode of treatment. The method according to claim 1,
The human subject is an adult. The method according to claim 1 or 2,
Said cancer is metastatic to the meninges. The method according to any one of claims 1 to 3,
The cancer metastatic to the CNS is a non-CNS solid tumor. The method according to claim 4,
Said solid tumor is selected from the group consisting of sarcoma, melanoma, ovarian cancer and rhabdomyosarcoma. The method according to claim 5,
Said solid tumor is selected from the group consisting of melanoma, ovarian cancer and rhabdomyosarcoma. The method according to any one of claims 1 to 6,
Said central nervous system (CNS) cancer is selected from the group consisting of neuroblastoma and primary recurrent CNS malignancy. The method according to any one of claims 1 to 7,
The antibody or antigen-binding fragment thereof is selected from the group consisting of murine antibodies or antigen-binding fragments thereof, humanized antibodies and antigen-binding fragments thereof, chimeric antibodies and antigen-binding fragments thereof, and human antibodies and antigen-binding fragments thereof. How to be. The method according to claim 8,
The antibody or antigen-binding fragment thereof is a murine antibody or antigen-binding fragment thereof. The method according to any one of claims 1 to 9,
The antibody or antigen-binding fragment thereof binds to the FG-loop of B7H3. The method according to any one of claims 1 to 10,
The antibody or antigen-binding fragment thereof
(a) a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3,
(b) a heavy chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 4,
(c) a heavy chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5,
(d) a light chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6,
(e) a light chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 7, and
(f) a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 8. The method according to any one of claims 1 to 11,
The antibody or antigen-binding fragment thereof
(a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1, and
(b) a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2. The method according to any one of claims 1 to 12,
The antibody or antigen-binding fragment thereof is administered intradurally to the subject. The method according to any one of claims 1 to 13,
The antibody or antigen-binding fragment thereof is administered to the subject via an intraventricular device. The method according to claim 14,
The ventricular device is an intraventricular catheter. The method according to claim 14,
The ventricular device is an intraventricular reservoir. The method according to any one of claims 1 to 16,
Wherein said radioisotope is ¹²⁴ I, ¹³¹ I, ¹⁷⁷ Lu, or ⁹⁹ mTc. The method according to any one of claims 1 to 17,
Administering said antibody or antigen-binding fragment thereof in one treatment cycle to said subject. The method according to any one of claims 1 to 18,
Administering the antibody or antigen-binding fragment thereof in two treatment cycles to the subject. The method according to claim 18 or 19,
One treatment cycle comprises a dosimetric dose and a therapeutic dose. The method according to any one of claims 1 to 20,
The therapeutically effective amount is about 10 mCi to about 200 mCi or about 10 mCi to about 100 mCi. The method according to any one of claims 1 to 21,
Said therapeutically effective amount is about 50 mCi. The method according to any one of claims 1 to 22,
The method extends the survival of the subject. The method according to any one of claims 1 to 23,
The method of extending sedation of the cancer in the subject. The method according to any one of claims 1 to 24,
Wherein said antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least about 80%, about 90%, about 95%, about 99% or about 100% homology with the amino acid sequence set forth in SEQ ID NO: 17. The method according to any one of claims 1 to 25,
The antibody or antigen-binding fragment thereof comprises the amino acid sequence set forth in SEQ ID NO: 17. The method according to any one of claims 1 to 26,
The antibody or antigen-binding fragment thereof has amino acids 224-241 of SEQ ID NO. The method according to any one of claims 1 to 26,
The antibody or antigen-binding fragment thereof has amino acids 242-267 of SEQ ID NO: 17. 17. The method according to any one of claims 1 to 28,
The treatment mode is selected from the group consisting of one or more chelator compounds, one or more chemotherapeutic agents, one or more checkpoint inhibitory agents, and radiation therapy. The method of claim 29,
The mode of treatment is a chelator compound. The method according to claim 29 or 30,
The antibody or antigen-binding fragment thereof is conjugated to a chelator compound, wherein the chelator compound is bound to a radioisotope. The method of any one of claims 29-31,
Said chelator compound is DOTA or DTPA. The method of claim 29,
The mode of treatment is monoclonal antibody 3F8 (MoAb 3F8), granulocyte-macrophage-colon-stimulating factor (GM-CSF), or a combination thereof. The method according to any one of claims 1 to 33,
Wherein said mode of treatment is administered systemically into and / or to said subject's CNS. The method according to any one of claims 1 to 34,
Wherein said mode of treatment is administered to said subject simultaneously or sequentially with said antibody or antigen-binding fragment thereof. An antibody or antigen-binding fragment thereof that specifically binds human B7H3,
The antibody or antigen-binding fragment thereof is conjugated to a chelator compound, wherein the chelator compound is bound to a radioisotope. The method of claim 36,
The antibody or antigen-binding fragment thereof is selected from the group consisting of murine antibodies and antigen-binding fragments thereof, humanized antibodies and antigen-binding fragments thereof, chimeric antibodies and antigen-binding fragments thereof, and human antibodies and antigen-binding fragments thereof. Antibody or antigen-binding fragment thereof. The method according to claim 36 or 37,
The antibody or antigen-binding fragment thereof is a murine antibody or antigen-binding fragment thereof. The method according to any one of claims 36 to 38,
The antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof that binds to the FG-loop of B7H3. The method according to any one of claims 36 to 39,
The antibody or antigen-binding fragment thereof
a. A heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 3,
b. A heavy chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 4,
c. A heavy chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 5,
d. Light chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 6,
e. A light chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 7, and
f. An antibody or antigen-binding fragment thereof comprising a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 8. The method according to any one of claims 36 to 40,
The antibody or antigen-binding fragment thereof
(a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1, and
(b) an antibody or antigen-binding fragment thereof comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2. The compound according to any one of claims 36 to 41, wherein
The radioisotope is ¹²⁴ I, ¹³¹ I, ¹⁷⁷ Lu, or ⁹⁹ mTc. Antibody or antigen-binding fragment thereof. The compound according to any one of claims 36 to 42, wherein
Said chelator compound is DOTA or DTPA antibody or antigen-binding fragment thereof. The method according to any one of claims 36 to 43,
The antibody or antigen-binding fragment thereof comprises an antibody or antigen thereof comprising an amino acid sequence having at least about 80%, about 90%, about 95%, about 99% or about 100% homology with the amino acid sequence set forth in SEQ ID NO: 17 Binding fragments. The method according to any one of claims 36 to 44,
The antibody or antigen-binding fragment thereof comprises an antibody or antigen-binding fragment thereof comprising the amino acid sequence shown in SEQ ID NO: 17. The method according to any one of claims 36 to 45,
The antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof having amino acids 224-241 of SEQ ID NO. The method according to any one of claims 36 to 46,
The antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof having amino acids 242-267 of SEQ ID NO. The method according to any one of claims 36 to 47,
The antibody or antigen-binding fragment thereof is a DOTA-8H9 conjugate or DTPA-8H9 conjugate. 49. The method of any of claims 36 to 48,
The antibody or antigen-binding fragment thereof is a ¹⁷⁷ Lu-DOTA-8H9 conjugate or a ¹⁷⁷ Lu-DTPA-8H9 conjugate or (177) LU-CHX-A "-DTPA-8H9, or an antigen-binding fragment thereof. The method according to any one of claims 36 to 49,
The antigen-binding fragment is a single chain variable fragment (scFv) antibody or antigen-binding fragment thereof. The method of claim 50,
The scFv is an antibody or antigen-binding fragment thereof comprising a portion of the amino acid sequence shown in SEQ ID NO: 9, SEQ ID NO: 13 and SEQ ID NO: 14. 52. A composition comprising the antibody of any one of claims 36-51 or an antigen-binding fragment thereof. A pharmaceutical composition comprising the antibody of claim 36 or an antigen-binding fragment thereof and a pharmaceutically acceptable carrier. The method of imaging a tumor in a subject, comprising administering to the subject the antibody of claim 36 or the antigen-binding fragment thereof. The compound according to any one of claims 36 to 51,
An antibody or antigen-binding fragment thereof for use as a medicament. The compound according to any one of claims 36 to 51,
Antibodies or antigen-binding fragments thereof for use in the treatment of cancer. The compound according to any one of claims 36 to 51,
An antibody or antigen-binding fragment thereof for use in the treatment of central nervous system (CNS) cancer. The compound according to any one of claims 36 to 51,
An antibody or antigen-binding fragment thereof for use in the treatment of metastatic CNS neuroblastoma, sarcoma, melanoma, ovarian carcinoma and primary recurrent CNS malignancy. The compound according to any one of claims 36 to 51,
An antibody or antigen-binding fragment thereof for use in a method of imaging a tumor in a subject. The compound according to any one of claims 36 to 51,
An antibody or antigen-binding fragment thereof for use in a method according to any one of claims 1 to 35. Use of the antibody or antigen-binding fragment thereof of any one of claims 36-51 for the manufacture of a medicament for killing and / or reducing tumor cells and / or inhibiting growth of tumors. Use of the antibody or antigen-binding fragment thereof of any one of claims 36-51 for preparing a medicament for imaging a tumor cell carrying an antigen recognized by the antibody or antigen-binding fragment thereof. Use of the antibody of any one of claims 36 to 51 or an antigen-binding fragment thereof for the manufacture of a medicament for a method according to any one of claims 1 to 35.

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